Method of treating acute respiratory distress syndrome (ARDS) or acute lung injury (ALI) associate with COVID-19 by administering an anti-LIGHT antibody

ABSTRACT

The present disclosure relates to methods of detecting free (active) LIGHT in biological samples to diagnose conditions associated with elevated free LIGHT, as well as to predict the effectiveness of anti-LIGHT therapies. The disclosure also relates to treating such conditions with anti-LIGHT antibodies. Conditions include acute lung injury (ALI) and acute respiratory distress syndrome (ARDS), optionally wherein the ALI and ARDS are associated with viral infection, including coronavirus infection. Conditions also include Crohn&#39;s Disease or an inflammatory condition associated with Crohn&#39;s Disease.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of priority to U.S. ProvisionalApplication No. 63/003,826, filed Apr. 1, 2020, U.S. ProvisionalApplication No. 63/027,127, filed May 19, 2020, and U.S. ProvisionalApplication No. 63/133,636, filed Jan. 4, 2021, the contents of each ofwhich are incorporated herein by reference for all purposes.

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Mar. 26, 2021, isnamed 01118-0047-00PCT_US_ST25.txt and is 27,637 bytes in size.

FIELD OF THE INVENTION

The present disclosure relates to methods of diagnosing and treatingsubjects with conditions associated with elevated free LIGHT levels,including subjects with Crohn's Disease (CD) or an inflammatorycondition associated with Crohn's Disease, subjects with immunedysregulation that may lead to multisystem organ failure, or subjectswith acute lung injury (ALI) or acute respiratory distress syndrome(ARDS), including those associated with coronavirus infection, includingCOVID-19. For example, in some embodiments, the subjects may be treatedwith anti-LIGHT antibodies. The disclosure also relates to a novel assayfor detecting free LIGHT.

BACKGROUND OF THE INVENTION

In December 2019, the spread of 2019 novel coronavirus 2019-nCoV(SARS-CoV-2) has emerged as a global emergency, causing both highmorbidity and mortality. SARS-CoV-2 originated in Wuhan China (Wu, F. etal. Nature 579, 265-269 (2020)) but has rapidly spread worldwide and hasbeen designated a global pandemic by the World Health Organization. Thevirus is highly infectious and it is estimated that up to 60% of theworld's population may eventually become infected by SARS-CoV-2. In theUS alone this would represent more than 180 million individuals.

COVID-19 is a disease caused by SARS-CoV-2. The initial presentation ofCOVID-19 infection includes fever with or without respiratory symptomsincluding cough, shortness of breath, and pneumonia. In most subjectsthe illness is mild and self-limited, however 15% to 20% experiencesevere respiratory illness, requiring hospitalization and oxygentherapy. (Huang, C. et al. Lancet 395, 497-506 (2020)). Many of thesesubjects require intensive care and ventilation owing to emergence ofacute respiratory distress syndrome (ARDS), (id.; Graham, R. L.,Donaldson, E. F. & Baric, R. S. Nature reviews. Microbiology 11, 836-848(2013)), which is a well-described and potentially fatal complication ofother viral respiratory syndromes (i.e., SARS, MERS, and H1N1). Othercomplications of COVID-19 include arrhythmia, shock, acute kidneyinjury, acute cardiac injury, liver dysfunction, and secondaryinfection. (Huang C, et al., Lancet (2020); Wang, D. et al., JAMA(2020)).

Accumulating evidence suggest that the main cause for mortality isunleashed immune response causing cytokine storm, acute lung injury, andAcute Respiratory Disease Syndrome (ARDS) resulting in fatal respiratoryfailure. Even in subjects who recover there may be long-lasting anddebilitating sequelae. There is an urgent need for cytokine-neutralizingtherapeutic agents, which will control COVID-19 associatedhyper-inflammation and ARDS.

In COVID-19 and other human corona respiratory virus (hCoV) infections,ARDS appears to result from a dysregulated hyperinflammatory responsemanifested by the release of excessive pro-inflammatory cytokines andchemokines, coined “cytokine storm.” (Channappanavar, R. & Perlman, S.Seminars in immunopathology 39, 529-539, (2017); Mehta, P. The Lancet(2020)). Cytokines and chemokines have long been thought to play animportant role in immunity and immunopathology during virus infections.A rapid and well-coordinated innate immune response is the first line ofdefense against viral infections, but dysregulated and excessive immuneresponses may cause immunopathology. (Fehr, A. R., Channappanavar, R. &Perlman, S. Annual review of medicine 68, 387-399 (2017);Channappanavar, R. et al. Cell host & microbe 19, 181-193 (2016)).Although there is no direct evidence for the involvement ofpro-inflammatory cytokines and chemokines in lung pathology during SARSand MERS, correlative evidence from subjects with severe diseasesuggests a role for hyper-inflammatory responses in hCoV pathogenesis.(Channappanavar, Seminars in immunopathology 39, 529-539 (2017); Mehta(2020); Sandoval-Montes, C. & Santos-Argumedo, L. Journal of leukocytebiology 77, 513-521 (2005); Xu et al., Microbiol 6(10):130 (2019)).

The cytokine storm in COVID-19 infection is thought to result frominitial rapid virus replication which may be more likely inimmunocompromised subjects. A notable feature of pathogenic humancoronaviruses such as SARS-CoV and MERS-CoV is that both virusesreplicate to high titers very early after infection both in vitro and invivo (Gralinski, L. E. & Baric, R. S. The Journal of pathology 235,185-195 (2015)). This high replication could lead to enhanced cytopathiceffects and production of higher levels of pro-inflammatory cytokinesand chemokines by infected epithelial cells. (Xiao, F. et al.Gastroenterology (2020)). These cytokines and chemokines in turnorchestrate massive infiltration of inflammatory cells into the lungs.(Gralinski, L. E. & Baric, R. S. The Journal of pathology 235, 185-195(2015)). Studies from hCoV infections in humans and experimental animalsdemonstrated a strong correlation between high SARS-CoV and MERS-CoVtiters and disease severity. Infection also appears to increasesecretion of cytokines (e.g., IL4 and IL10) which in turn can increase Tcell activation. (Sandoval-Montes (2005); Xu, Z. et al. The Lancet.Respiratory medicine (2020)). Thus, the cytokine storm that drivestissue injury and vascular permeability in the lungs is likely mediated,in part by T cell activation with increased expression of cytokines.

In addition, reports indicate that pulmonary (lung) fibrosis, which isknown to be a result of ARDS, is a known COVID-19 infectioncomplication. (Huang, C. et al. Lancet 395, 497-506 (2020)).

Both human and animal studies demonstrate accumulation of inflammatorymonocyte-macrophages and neutrophils in the lungs following hCoVinfection. These cells are the predominant source of cytokines andchemokines associated with hCoV lethal disease observed both in humansand animal models. (Channappanavar, Seminars in immunopathology (2017)).

While a primary focus of treatment of COVID-19 is the development ofappropriate antiviral and vaccination approaches, currently noestablished therapy exists for treatment of ARDS associated withCOVID-19. Agents targeting cytokine storm have includedcytokine-directed therapies, including IL-1β and IL-6 antagonists;however, there is no established single therapy for the treatment and/orprevention of ALI associated with cytokine storm. The development of asafe and effective therapy for COVID-19-associated acute lung injury(ALI) and ARDS could significantly reduce the mortality and postinfectious morbidity of this global pandemic and alleviate the severestrain placed on healthcare systems.

Further, the initial clinical sign of COVID-19 that allowed casedetection was pneumonia (Chan, J F, et al., Lancet (2020)).Complications of COVID-19 pneumonia include acute respiratory distresssyndrome (ARDS), arrhythmia, shock, acute kidney injury, acute cardiacinjury, liver dysfunction, and secondary infection (Huang C, et al.,Lancet (2020); Wang, D. et al., JAMA (2020)). The main cause ofmortality in COVID-19 appears to be dysregulated hyperimmune responsecausing cytokine storm, acute lung injury and ARDS. Fifteen to 20% ofCOVID-19 patients experience severe respiratory illness, requiringhospitalization and oxygen therapy (Huang C, et al., Lancet (2020)).There are currently no treatments to prevent progression of COVID-19pneumonia to ARDS in patients with COVID-19.

Crohn's Disease (CD) is an idiopathic, chronic, inflammatory conditionof the gastrointestinal tract with a high risk for complications andneed for surgical interventions. Crohn's Disease is a life-long disorderthat may become clinically apparent at almost any time from earlychildhood to late adulthood. (Freeman, Natural history and long-termclinical course of Crohn's disease, World J. Gastroenterol., 2014:20(1);31-36). The typical age of detection or diagnosis of the disease isusually during the late teens and early twenties, and during the lasttwo to three decades, over 80% of patients with Crohn's Disease arediagnosed before age 40. (Freeman 2014).

Crohn's Disease may impact the entire gastrointestinal tract. (Shi andNg, The state of the art on treatment of Crohn's disease, J.Gastroenterol., 2018:53; 989-998). The majority of patients have achronic intermittent course during 10 years after diagnosis. The diseaseappears to be progressive, although the rate of progression may bealtered or slowed by the use of medication or with surgicalintervention. (Freeman 2014). Common symptoms include diarrhea,abdominal pain, rectal bleeding, fever, weight loss, and fatigue.(Veauthier and Hornecker, Crohn's Disease: Diagnosis and Management, Am.Fam. Physician, 2018:98(11); 661-669).

Corticosteroids and thiopurines remain the main treatments, whileanti-TNF agents are being increasingly prescribed earlier in diseasecourse. (Shi and Ng 2018). Anti-TNF therapies are recommended inpatients with high risk for unfavorable prognosis. (Shi and Ng 2018).However, primary non-response or secondary loss of response to anti-TNFtherapy occurs in a large proportion of patients. (Shi and Ng 2018).Therefore, new and improved therapies are needed.

An important immunoregulatory cytokine, LIGHT (acronym for “homologousto Lymphotoxin, exhibits Inducible expression and competes with HSVGlycoprotein D for binding to HVEM (herpesvirus entry mediator), areceptor expressed on T lymphocytes”), also known as TNFSF14 (tumornecrosis factor superfamily member 14) is secreted in high levels duringviral infection, which supports ARDS-related pulmonary fibrosis andcytokine storm. (Xu, W. et al. Frontiers in Microbiology 10, 130(2019)). Neutrophils and macrophages express high levels of LIGHT andTNF and are a major source of these inflammatory cytokines. (Kwon, B. S.et al. The Journal of Biological Chemistry 272, 14272-14276 (1997)).

LIGHT (TNFSF 14) belongs to the tumor necrosis factor superfamily and isexpressed by activated T cells, monocytes-macrophages and additionaltypes of antigen presenting cells. LIGHT is considered one of the“Master Regulators” of the immune system and has a key role in thecommunication system which controls immune response. LIGHT has a dualmechanism of action; exerting its effects by activating both T cells andB cells as well as upregulating other inflammatory cytokines.

LIGHT activates two key receptors, herpesvirus entry mediator (HVEM) andlymphotoxin β receptor (LTβR), both expressed on lung epithelial cells.Early in infection LIGHT released from neutrophils and macrophages bindcellular receptors, which causes inflammatory cell infiltration,releasing high level of TNF and additional pro-inflammatory cytokines.LIGHT also has a co-stimulatory role in T cell activation drivingproinflammatory and tissue damaging effects. (Ware, C. F. Advances inexperimental medicine and biology 647, 146-155 (2009); Ware, C. F.Immunological reviews 223, 186-201 (2008)). Therefore, LIGHT has rolesin many immune-mediated pathologies such as Crohn's Disease, IBD,Rheumatoid arthritis, and fibrosis. An additional receptor for LIGHT isa decoy receptor (coined DCR3), which binds LIGHT and interferes withits activity by competing with receptor binding. (Steinberg, M. W., etal., M. Seminars in immunopathology 31, 207-221 (2009); Wroblewski, V.J. et al. Biochemical pharmacology 65, 657-667 (2003)). In hyperinflammation and cytokine storm conditions, DcR3 is likely to beoverwhelmed, generating high DCR3-free (active) LIGHT.

LIGHT has been shown to play a key role in viral pneumonia. LIGHTprotein has been reported to be elevated in PBMCs of subjects presentingsevere pneumonia caused by viral infection, as a part of the TNF familyand IL-1 family of genes and cognate proteins elevated in these subjectscompared to healthy controls. (Xu (2019)). LIGHT levels correlate withdisease severity—as disease progressed from minor to severe, LIGHTlevels were elevated.

High LIGHT levels in the lung may be a driver of pulmonary fibrosis.Alveolar and interstitial fibrosis is a hallmark of ARDS, (Marshall, R.,Bellingan, G. & Laurent, G. Thorax 53, 815-817 (1998)) and is a causefor further lung injury and the need for supportive mechanicalventilation. Over-activated fibroblasts are a major cause for pulmonaryfibrosis. After an infection, fibroblasts proliferate, differentiateinto myofibroblasts and migrate to the alveolar airspace. Over activatedmyofibroblasts secrete extra cellular matrix and form attachments to thebasement membrane. Consequently, this process results in obliteration ofalveolar spaces with an irregular extracellular matrix. (Quesnel, C. etal. The European respiratory journal 35, 1312-1321 (2010)). LIGHTsupports pulmonary fibrosis in several mechanisms. Recently, da Silva etal (da Silva Antunes, R., Mehta, A. K., Madge, L., Tocker, J. & Croft,M. Front Immunol 9, 576 (2018)), described the role of LIGHT via itsreceptor LTßR in promoting fibroblasts proliferation in the process ofpulmonary fibrosis.

Genetic deficiency in LIGHT, and blocking LIGHT binding to both of itsreceptors, strongly reduced tissue remodeling and fibrosis in the lungsof allergen-challenged mice in models of severe asthma and in a model ofidiopathic pulmonary fibrosis. (Doherty, T. A. et al. Nature medicine17, 596-603, (2011)). Neutralizing LIGHT demonstrate reduced fibrosisphenotype. (Da Silva (2018); Herro, R. & Croft, M. Pharmacol Res 104,151-155(2016)). This strategy of neutralizing LIGHT as a treatment forfibrosis should be relevant for ARDS derived fibrosis. High LIGHT levelsinduce high cytokines secretion by bronchial and alveolar epithelialcells in-vitro, via LTßR and HVEM (TNFRSF14), which supportsteroid-resistant lung inflammation. (da Silva Antunes, R., Madge, L.,Soroosh, P., Tocker, J. & Croft, M. Journal of immunology (Baltimore,Md.: 1950) 195, 2429-2441(2015); Herro, R., Da Silva Antunes, R.,Aguilera, A. R., Tamada, K. & Croft, M. The Journal of allergy andclinical immunology 136, 757-768 (2015)).

LIGHT has a role as an important mediator in mucosal inflammation andinflammatory bowel disease (IBD) pathogenesis (Cohavy et al., LIGHTexpression by mucosal T cells may regulate IFN-gamma expression in theintestine, J. Immunol., 2004; 173(1):251-8; Ware, C F, NetworkCommunications: Lymphotoxins, LIGHT and TNF, Annual Rev. Immunol., 2005:23:787-819; Cohavy et al., LIGHT is constitutively expressed on T and NKcells in the human gut and can be induced by CD2-mediated signaling, J.Immunol., 2005:174:646-53; Wang et al., The critical role of LIGHT inpromoting intestinal inflammation and Crohn's disease, J. Immunol.,2005:174; 8173-82). The human LIGHT gene maps to chromosome 19p13.3, aregion that has been implicated in the pathogenesis of CD (Granger etal., Genomic characterization of light reveals linkage to an immuneresponse locus on chromosome 19p13.3 and distinct isoforms generated byalternate splicing or proteolysis, J Immunol., 2001:167:5122-28; Riouxet al., Genome wide search in Canadian families with inflammatory boweldisease reveals two novel susceptibility loci, Am. J. Hum. Genet.,2000:66:1863-70). The concept that LIGHT provides a criticalpro-inflammatory signal during cellular immune responses is reinforcedby studies in IBD patients. LIGHT messenger ribonucleic acid (RNA) isupregulated in biopsies from inflamed areas of small bowel (Cohavy etal., LIGHT is constitutively expressed on T and NK cells in the humangut and can be induced by CD2-mediated signaling, J. Immunol.,2005:174:646-53). Decoy receptor 3 belongs to the TNF superfamily(TNFRSF6B) (Yu et al., A newly identified member of tumor necrosisfactor receptor superfamily (TR6) suppresses LIGHT-mediated apoptosis,J. Biol. Chem., 1999: 274(20):13733-6). It acts as a decoy receptor thatcompetes with death receptors for ligand binding and is postulated toplay a regulatory role in suppressing Fas ligand (FasL)- andLIGHT-mediated cell death and T cell activation as well as to induceangiogenesis via neutralization of TNF-like ligand 1A (TL1A) (Yu et al.1999). Decoy receptor 3 is over-expressed in the epithelial layer ofileum specimens in patients with CD, both at actively inflamed andnon-active sites. Decoy receptor 3 serum levels are significantlyelevated in patients with active and non-active CD compared with healthycontrols. The expression of DcR3 in intestinal epithelial cells isinduced by TNFα. Increased DcR3 expression is associated with activationof nuclear factor kappa B (NF-κB) and results in protection ofintestinal epithelial cells and lamina propria T cells fromCD95L-induced apoptosis (Funke et al., Functional characterisation ofdecoy receptor 3 in Crohn's disease. Gut, 2009: April; 58(4):483-91).Defective variants of DcR3 have recently been observed in patients withpediatric onset IBD which further suggests an important protective rolefor DcR3 (Cardinale et al., Targeted resequencing identifies defectivevariants of decoy receptor 3 in pediatric-onset inflammatory boweldisease, Genes Immun. 2013: Oct.; 14(7):447-52), potentially bymoderating the effects of TNF and LIGHT.

The roles of LIGHT and DcR3 in the pathogenesis of IBD, described above,provide a rationale for the study of an anti-LIGHT monoclonal antibodyin CD patients with or without loss-of-function mutations in DcR3.

Most currently available assays only measure total LIGHT, which includesLIGHT bound to its receptors, including DcR3. Total LIGHT may notprovide as accurate of a picture of the levels of LIGHT causing disease,which may be free, unbound LIGHT. Thus, there is a need for improvedLIGHT assays that measure free LIGHT alone.

SUMMARY OF THE INVENTION

The present disclosure includes, for example, any one or a combinationof the following embodiments:

-   Embodiment 1. A method of detecting the presence of free LIGHT in a    biological sample of a subject comprising the steps of:    -   (a) contacting the biological sample with at least one        anti-LIGHT antibody;    -   (b) incubating the biological sample to allow the anti-LIGHT        antibody to bind to free LIGHT; and    -   (c) detecting the presence of complexes formed between the        anti-LIGHT antibody and free LIGHT in the biological sample.-   Embodiment 2. A method of diagnosing a condition associated with    elevated free LIGHT in a subject comprising the steps of:    -   (a) contacting a biological sample with at least one anti-LIGHT        antibody;    -   (b) incubating the biological sample to allow the anti-LIGHT        antibody to bind to free LIGHT;    -   (c) detecting the presence of complexes formed between the        anti-LIGHT antibody and free LIGHT in the biological sample; and    -   (d) diagnosing the subject as having a condition associated with        elevated free LIGHT if a higher level of free LIGHT is detected        as compared to a control.-   Embodiment 3. A method of treating a condition associated with    elevated free LIGHT, comprising administering to a subject in need    thereof an effective amount of an anti-LIGHT antibody.-   Embodiment 4. A method of treating a condition associated with    elevated free LIGHT in a subject in need thereof, comprising:    -   (a) contacting a biological sample isolated from the subject        with a first anti-LIGHT antibody;    -   (b) incubating the biological sample to allow the first        anti-LIGHT antibody to bind to free LIGHT;    -   (c) detecting the presence of complexes formed between the first        anti-LIGHT antibody and free LIGHT in the biological sample; and    -   (d) administering to the subject an effective amount of a second        anti-LIGHT antibody, wherein the first and the second antibody        differ, thereby treating the condition associated with elevated        free LIGHT.-   Embodiment 5. The method of any one of embodiments 2-4, wherein the    condition associated with elevated free LIGHT comprises any one or    more of:    -   (a) inflammation, optionally wherein the inflammation is        hyperinflammation;    -   (b) immune dysregulation that leads to multisystem organ        failure;    -   (c) acute lung injury (ALI), optionally wherein the ALI is        associated with a bacterial or viral infection, including        coronavirus infection;    -   (d) acute respiratory distress syndrome (ARDS), optionally        wherein the ARDS is associated with a bacterial or viral        infection, including coronavirus infection;    -   (e) cytokine storm that drives tissue injury and vascular        permeability;    -   (f) post-infection pulmonary fibrosis; and    -   (g) pneumonia, optionally wherein the pneumonia is associated        with a bacterial or viral infection, including coronavirus        infection.-   Embodiment 6. The method of any one of embodiments 2, 4, and 5,    wherein the detection of free LIGHT indicates that treatment of the    condition associated with elevated free LIGHT with an anti-LIGHT    antibody will be effective.-   Embodiment 7. The method of any one of embodiments 3-6, wherein the    anti-LIGHT antibody administered to the subject suppresses T cell    activation.-   Embodiment 8. The method of any one of embodiments 3-7, wherein the    anti-LIGHT antibody administered to the subject suppresses increased    expression of cytokines.-   Embodiment 9. The method of any one of embodiments 3-8, wherein the    anti-LIGHT antibody administered to the subject reduces the    subject's risk of mortality or morbidity.-   Embodiment 10. The method of any one of embodiments 3-9, wherein the    anti-LIGHT antibody administered to the subject prevents progression    to ARDS.-   Embodiment 11. The method of any one of embodiments 3-10, wherein    the anti-LIGHT antibody administered to the subject prevents the    need for ventilation/intubation of the subject.-   Embodiment 12. A method of treating severe COVID-19 pneumonia    comprising administering an anti-LIGHT antibody to a subject in need    thereof.-   Embodiment 13. A method of treating acute inflammatory disease    associated with COVID-19 pneumonia comprising administering an    anti-LIGHT antibody to a subject in need thereof.-   Embodiment 14. A method of treating respiratory failure associated    with COVID-19 pneumonia comprising administering an anti-LIGHT    antibody to a subject in need thereof.-   Embodiment 15. A method of treating cytokine storm comprising    administering an anti-LIGHT antibody to a subject in need thereof.-   Embodiment 16. A method of treating a dysregulated hyperimmune    response (sometimes referred to as “cytokine storm”) comprising    administering an anti-LIGHT antibody to a subject in need thereof.-   Embodiment 17. A method of treating Acute Respiratory Disease    Syndrome (ARDS) comprising administering an anti-LIGHT antibody to a    subject in need thereof.-   Embodiment 18. The method of any one of embodiments 1 and 2 wherein    the anti-LIGHT antibody comprises a heavy chain and a light chain    that together comprise one of the following sets of CDR-H1, CDR-H2,    CDR-H3, CDR-L1, CDR-L2, and CDR-L3 amino acid sequences:    -   (a) SEQ ID NOs: 10, 11, 12, 13, 14, and 15;    -   (b) SEQ ID NOs: 16, 17, 18, 19, 20, and 21;    -   (c) SEQ ID NOs: 22, 23, 24, 25, 26, and 27;    -   (d) SEQ ID NOs: 28, 29, 30, 31, 32, and 33;    -   (e) SEQ ID NOs: 34, 35, 36, 37, 38, and 39;    -   (f) SEQ ID NOs: 40, 41, 42, 43, 44, and 45;    -   (g) SEQ ID NOs: 46, 47, 48, 49, 50, and 51; and    -   (h) SEQ ID NOs: 52, 53, 54, 55, 56, and 57.-   Embodiment 19. The method of embodiment 4, wherein the first    anti-LIGHT antibody comprises a heavy chain and a light chain that    together comprise one of the following sets of CDR-H1, CDR-H2,    CDR-H3, CDR-L1, CDR-L2, and CDR-L3 amino acid sequences:    -   (a) SEQ ID NOs: 10, 11, 12, 13, 14, and 15;    -   (b) SEQ ID NOs: 16, 17, 18, 19, 20, and 21;    -   (c) SEQ ID NOs: 22, 23, 24, 25, 26, and 27;    -   (d) SEQ ID NOs: 28, 29, 30, 31, 32, and 33;    -   (e) SEQ ID NOs: 34, 35, 36, 37, 38, and 39;    -   (f) SEQ ID NOs: 40, 41, 42, 43, 44, and 45;    -   (g) SEQ ID NOs: 46, 47, 48, 49, 50, and 51; and    -   (h) SEQ ID NOs: 52, 53, 54, 55, 56, and 57,        and wherein the second anti-LIGHT antibody comprises a heavy        chain and a light chain that together comprise any one of the        sets of CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 of        (a)-(h) above or SEQ ID NOs: 2, 3, 4, 5, 6, and 7.-   Embodiment 20. The method of any one of embodiments 3 and 5-17,    wherein the anti-LIGHT antibody that is administered to the subject    comprises a heavy chain and a light chain that together comprise one    of the following sets of CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and    CDR-L3 amino acid sequences:    -   (a) SEQ ID NOs: 2, 3, 4, 5, 6, and 7;    -   (b) SEQ ID NOs: 10, 11, 12, 13, 14, and 15;    -   (c) SEQ ID NOs: 16, 17, 18, 19, 20, and 21;    -   (d) SEQ ID NOs: 22, 23, 24, 25, 26, and 27;    -   (e) SEQ ID NOs: 28, 29, 30, 31, 32, and 33;    -   (f) SEQ ID NOs: 34, 35, 36, 37, 38, and 39;    -   (g) SEQ ID NOs: 40, 41, 42, 43, 44, and 45;    -   (h) SEQ ID NOs: 46, 47, 48, 49, 50, and 51; and    -   (i) SEQ ID NOs: 52, 53, 54, 55, 56, and 57.-   Embodiment 21. The method of any one of the preceding embodiments,    wherein the antibody comprises the CDR-H1, CDR-H2, CDR-H3, CDR-L1,    CDR-L2, and CDR-L3 amino acid sequences of any one of antibodies    1C02, 13H04, 31A10, 1C06, 98C07, 18E04, 42A02, 29C09, 14B09, 117C06,    114F05, or 62C01 described in WO 2015/107331.-   Embodiment 22. The method of any one of embodiments 2-21, wherein    the condition associated with elevated free LIGHT is coronavirus    infection.-   Embodiment 23. The method of embodiment 22, wherein the condition    associated with elevated free LIGHT is a COVID-19 infection.-   Embodiment 24. The method of embodiment 23, wherein the coronavirus    infection is a MERS-CoV or SARS-CoV infection.-   Embodiment 25. The method of any one of embodiments 2-24, wherein    the condition associated with elevated free LIGHT is Crohn's Disease    or an inflammatory condition associated with Crohn's Disease.-   Embodiment 26. The method of any one of the preceding embodiments,    wherein the anti-LIGHT antibody administered comprises a heavy chain    and a light chain comprising a CDR-H1, CDR-H2, CDR-H3, CDR-L1,    CDR-L2, and CDR-L3 amino acid sequences of SEQ ID NOs: 2, 3, 4, 5,    6, and 7, respectively.-   Embodiment 27. The method of any one of the preceding embodiments,    wherein a single dose of about 16 mg/kg of the anti-LIGHT antibody    is administered.-   Embodiment 28. The method of any one of the preceding embodiments,    wherein the subject has received, or is currently receiving, an    anti-COVID-19 therapy, optionally wherein the therapy is    corticosteroids, hydroxychloroquine, and/or remdesivir.-   Embodiment 29. The method of embodiment 28, wherein the dose of    corticosteroid is considered high dose.-   Embodiment 30. The method of any one of the preceding embodiments,    wherein the subject is human.-   Embodiment 31. The method of any one of the preceding embodiments,    wherein the subject has a respiratory disease, optionally caused by    a coronavirus infection.-   Embodiment 32. The method of any one of the preceding embodiments,    wherein the subject has pneumonia.-   Embodiment 33. The method of any one of the preceding embodiments,    wherein the subject has acute lung injury (ALI).-   Embodiment 34. The method of any one of the preceding embodiments,    wherein the subject has acute respiratory distress syndrome (ARDS).-   Embodiment 35. The method of any one of the preceding embodiments,    wherein the subject has a mild coronavirus infection.-   Embodiment 36. The method of any one of the preceding embodiments,    wherein the subject has a moderate coronavirus infection.-   Embodiment 37. The method of any one of the preceding embodiments,    wherein the subject has a severe coronavirus infection.-   Embodiment 38. The method of any one of the preceding embodiments,    wherein the subject is at the Early Infection (Stage I) of a    coronavirus infection.-   Embodiment 39. The method of any one of the preceding embodiments,    wherein the subject is at the Pulmonary Phase (Stage II) of a    coronavirus infection.-   Embodiment 40. The method of any one of the preceding embodiments,    wherein the subject is at the Hyperinflammation Phase (Stage III) of    a coronavirus infection.-   Embodiment 41. The method of any one of the preceding embodiments,    wherein the subject is a pediatric subject.-   Embodiment 42. The method of any one of the preceding embodiments,    wherein the subject is an adult.-   Embodiment 43. A kit for use in a method of any one of embodiments    1-3, 5-18, and 20-42 comprising an anti-LIGHT antibody and reagents    for carrying out the method.-   Embodiment 44. A kit for use in a method of any one of embodiments 4    and 19 comprising a first anti-LIGHT antibody and a second-LIGHT    antibody, wherein the first and the second antibody differ, and    reagents for carrying out the method.-   Embodiment 45. The kit of embodiment 43, further comprising a solid    phase to which the anti-LIGHT antibody is attached.-   Embodiment 46. The kit of embodiment 44, further comprising a solid    phase to which the first anti-LIGHT antibody is attached.-   Embodiment 47. The kit of embodiment 43 or 44, further comprising a    solid phase to which free LIGHT derived from the biological sample    is attached.-   Embodiment 48. A method of determining the amount of free/non-bound    Tumor Necrosis Factor Superfamily member 14 (TNFSF14 or LIGHT) in a    sample suspected to contain free LIGHT from a subject comprising:    -   (a) contacting a sample with a capturing molecule for free LIGHT        that specifically binds to free LIGHT, but not to bound LIGHT;    -   (b) incubating the sample to allow the capturing molecule to        bind to free LIGHT;    -   (c) detecting the binding of free LIGHT to the capturing        molecule and determining the amount of free LIGHT in the sample.-   Embodiment 49. The method of embodiment 48, wherein the capturing    molecule is an antibody, optionally wherein the antibody comprises    an anti-LIGHT antibody comprising a heavy chain and a light chain    that together comprise one of the following sets of CDR-H1, CDR-H2,    CDR-H3, CDR-L1, CDR-L2, and CDR-L3 amino acid sequences:    -   (a) SEQ ID NOs: 10, 11, 12, 13, 14, and 15;    -   (b) SEQ ID NOs: 16, 17, 18, 19, 20, and 21;    -   (c) SEQ ID NOs: 22, 23, 24, 25, 26, and 27;    -   (d) SEQ ID NOs: 28, 29, 30, 31, 32, and 33;    -   (e) SEQ ID NOs: 34, 35, 36, 37, 38, and 39;    -   (f) SEQ ID NOs: 40, 41, 42, 43, 44, and 45;    -   (g) SEQ ID NOs: 46, 47, 48, 49, 50, and 51; and    -   (h) SEQ ID NOs: 52, 53, 54, 55, 56, and 57.-   Embodiment 50. The method of any one of embodiments 48-49, wherein    the capturing molecule specifically binds to free LIGHT, but not to    a LIGHT/DcR3 complex, or LIGHT/HVEM complex, or LIGHT/LTβR complex.-   Embodiment 51. The method of any one of embodiments 48-50, wherein    the capturing molecule specifically binds to free LIGHT at the site    at which LIGHT binds to DcR3 or in the vicinity of the site at which    LIGHT binds to DcR3.-   Embodiment 52. The method of embodiment 51, wherein the capturing    molecule specifically binds to free LIGHT at a site at which LIGHT    binds to DcR3.-   Embodiment 53. The method of any one of embodiments 48-52, wherein a    detection molecule is provided, wherein the detection molecule binds    to LIGHT at site that is different from the site at which the    capturing molecule binds.-   Embodiment 54. The method of any one of embodiments 48-53, further    comprising comparing the amount of free and total LIGHT in the    sample.-   Embodiment 55. The method of any one of embodiments 48-54, wherein    the capturing molecule is an antibody.-   Embodiment 56. The method of any one of embodiments 48-55, wherein    the capturing molecule is an antibody chosen from monoclonal,    polyclonal, chimeric, single chain, bispecific or bi-effective,    simianized, human and humanized antibodies.-   Embodiment 57. The method of any one of embodiments 48-56, wherein    the capturing antibody is monoclonal antibody.-   Embodiment 58. The method of any one of embodiments 48-57, wherein    the capturing antibody is bound to a support (e.g., nanoparticle in    Simoa platform).-   Embodiment 59. The method of any one of embodiments 48-58, wherein    the capturing antibody is Enzo ALX-804-841-C100.-   Embodiment 60. The method of any one of embodiments 48-59, wherein    the detection molecule is an antibody.-   Embodiment 61. The method of any one of embodiments 48-60, wherein    the detection molecule is monoclonal antibody.-   Embodiment 62. The method of any one of embodiments 48-61, wherein    the capturing antibody is Enzo ALX-804-841-C100 and the detection    antibody is ProSci RF16062.-   Embodiment 63. The method of any one of embodiments 48-62, wherein    said sample is serum, plasma, saliva, or stool.-   Embodiment 64. The method according to any one of embodiments 48-63,    wherein said sample is a serum sample.-   Embodiment 65. A method of treating Crohn's Disease or an    inflammatory condition associated with Crohn's Disease, comprising    administering an anti-LIGHT antibody to a subject in need thereof,    wherein the anti-LIGHT antibody comprises a heavy chain and a light    chain that together comprise one of the following sets of CDR-H1,    CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 amino acid sequences:    -   (a) SEQ ID NOs: 2, 3, 4, 5, 6, and 7;    -   (b) SEQ ID NOs: 10, 11, 12, 13, 14, and 15;    -   (c) SEQ ID NOs: 16, 17, 18, 19, 20, and 21;    -   (d) SEQ ID NOs: 22, 23, 24, 25, 26, and 27;    -   (e) SEQ ID NOs: 28, 29, 30, 31, 32, and 33;    -   (f) SEQ ID NOs: 34, 35, 36, 37, 38, and 39;    -   (g) SEQ ID NOs: 40, 41, 42, 43, 44, and 45;    -   (h) SEQ ID NOs: 46, 47, 48, 49, 50, and 51; and    -   (i) SEQ ID NOs: 52, 53, 54, 55, 56, and 57.-   Embodiment 66. The method of embodiment 25 or 65, wherein a dose of    1.0 mg/kg of the anti-LIGHT antibody is administered every 14 days.-   Embodiment 67. The method of embodiment 25 or 65, wherein a dose of    3.0 mg/kg of the anti-LIGHT antibody is administered every 14 days.-   Embodiment 68. The method of any one of embodiments 65-67, wherein    the subject is a human.-   Embodiment 69. The method of any one of embodiments 65-68, wherein    the subject is an adult.-   Embodiment 70. The method of any one of embodiments 65-69, wherein    the subject has failed treatment with an approved therapeutic dose    of an anti-TNFα monoclonal antibody treatment with either no initial    response or an initial response to induction with subsequent lost    response.-   Embodiment 71. The method of any one of embodiments 65-70, wherein    administration of the anti-LIGHT antibody reduces the subject's CDAI    score.-   Embodiment 72. The method of any one of embodiments 65-71, wherein    administration of the anti-LIGHT antibody decreases the subject's    SES-CD score.-   Embodiment 73. The method of any one of embodiments 65-72, wherein    administration of the anti-LIGHT antibody increases the subject's    IBD-Q score.-   Embodiment 74. The method of any one of embodiments 3-24 and 26-42,    wherein administration of the anti-LIGHT antibody reduces serum    free-LIGHT levels in the subject by 85% or more.-   Embodiment 75. The method of embodiment 74, wherein the reduction in    serum free-LIGHT levels occurred in less than 5 days after    administration of the anti-LIGHT antibody.-   Embodiment 76. The method of embodiment 74, wherein the reduction in    serum free-LIGHT levels occurred in about 1 day after administration    of the anti-LIGHT antibody.-   Embodiment 77. The method of any one of embodiments 3-24, 26-42, and    74-76 wherein administration of the anti-LIGHT antibody reduces the    subject's risk of mortality by equal to or greater than 50% at 60    days after administration.-   Embodiment 78. The method of any one of embodiments 3-24, 26-42, and    74-76, wherein administration of the anti-LIGHT antibody reduces the    subject's risk of mortality by equal to or greater than 50% at 28    days after administration.-   Embodiment 79. The method of any one of embodiments 3-24, 26-42, and    74-78, where the subject is 60 years of age or older.-   Embodiment 80. The method of embodiment 79, wherein administration    of the anti-LIGHT antibody shortens the length of the subject's    hospital stay compared to subjects receiving standard of care    treatment.-   Embodiment 81. The method of any one of embodiments 3-24, 26-42, and    74-80, wherein administration of the anti-LIGHT antibody reduces the    subject's risk of respiratory failure.-   Embodiment 82. The method of any one of embodiments 3-42, 65-73, and    77-81, wherein administration of the anti-LIGHT antibody reduces    serum free-LIGHT levels in the subject.-   Embodiment 83. Use of an anti-LIGHT antibody in the manufacture of a    medicament for treating a condition associated with elevated free    LIGHT.-   Embodiment 84. A composition comprising an anti-LIGHT antibody for    use as a medicament in the treatment of a condition associated with    elevated free LIGHT.-   Embodiment 85. A composition comprising an anti-LIGHT antibody for    use in treating a condition associated with elevated free LIGHT.-   Embodiment 86. The use or composition for use according to any one    of embodiments 81-83, wherein the condition associated with elevated    free LIGHT is one or more of:    -   a. inflammation, optionally wherein the inflammation is        hyperinflammation;    -   b. immune dysregulation that leads to multisystem organ failure;    -   c. acute lung injury (ALI), optionally wherein the ALI is        associated with a bacterial or viral infection, including        coronavirus infection;    -   d. acute respiratory distress syndrome (ARDS), optionally        wherein the ARDS is associated with a bacterial or viral        infection, including coronavirus infection;    -   e. cytokine storm that drives tissue injury and vascular        permeability;    -   f. post-infection pulmonary fibrosis;    -   g. pneumonia, optionally wherein the pneumonia is associated        with a bacterial or viral infection, including coronavirus        infection;    -   h. Crohn's Disease or an inflammatory condition associated with        Crohn's Disease; or    -   i. COVID-19 infection.-   Embodiment 87. The use or composition for use according to any one    of embodiments 83-86, wherein the anti-LIGHT antibody comprises a    heavy chain and a light chain that together comprise one of the    following sets of CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3    amino acid sequences:    -   (a) SEQ ID NOs: 2, 3, 4, 5, 6, and 7;    -   (b) SEQ ID NOs: 10, 11, 12, 13, 14, and 15;    -   (c) SEQ ID NOs: 16, 17, 18, 19, 20, and 21;    -   (d) SEQ ID NOs: 22, 23, 24, 25, 26, and 27;    -   (e) SEQ ID NOs: 28, 29, 30, 31, 32, and 33;    -   (f) SEQ ID NOs: 34, 35, 36, 37, 38, and 39;    -   (g) SEQ ID NOs: 40, 41, 42, 43, 44, and 45;    -   (h) SEQ ID NOs: 46, 47, 48, 49, 50, and 51; and    -   (i) SEQ ID NOs: 52, 53, 54, 55, 56, and 57.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows capture of free LIGHT (i.e. DCR-free LIGHT, active LIGHT)with candidate antibody pair (capture antibody: Enzo ALX-804-841-C100,detection antibody: ProSci RF16062; specific epitopes are not detailedfor these antibodies), with linearity conducted at 1:10, 1:20, and 1:40dilution. Simoa™ ultra-high sensitive assay (Myriad RBM) was used todetect and measure free LIGHT with high sensitivity, using Quanterix'sfully automated immunoassay platform: Simoa HD-1 Analyzer and singlemolecule array (Simoa) technology. All incubations take place at roomtemperature inside the Simoa HD-1 analyzer. Capture antibody conjugatedparamagnetic beads were incubated with standards, samples or controlsand biotinylated detection antibodies. The beads were then washed andincubated with streptavidin-ß-galactosidase (SßG). After the final wash,the beads were loaded into the Simoa Disc with enzyme substrate,resorufin ß-galactopyranoside (RGP). The fluorescence signals arecompared to the standard curve and the quantity of LIGHT Free isdetermined for each sample. After screening anti-LIGHT antibodies inpairs for sandwich immunoassay-based detection of free LIGHT, assayswith one candidate pair were performed to test for linearity andspecificity.

FIG. 2A-B shows capture of free LIGHT with candidate pair, withlinearity conducted at 1:10, 1:20, 1:40, 1:80 dilution, shown as a tablein FIG. 2A and as a graph in FIG. 2B. The same assay described in FIG. 1was performed at different dilutions.

FIG. 3 shows DcR3 interference of the capture of free LIGHT was testedon the free LIGHT detecting candidate pair (Enzo ALX-804-841-C100-ProSciRF16062 (capture-detection)). A diluent containing free LIGHT was used,rather than native free LIGHT in a serum or plasma sample. DcR3 spikedconcentration was 10,000 ng/ml and 11 additional lower concentrations.Signal inhibition value was calculated as a signal reduction (MFI) forthe Enzo/ProSci pair.

FIG. 4A-C show comparison of DcR3 interference of free LIGHT detectingantibody pairs in spiked and unspiked samples. FIG. 4A: A spike andrecovery experiment was performed to assess DcR3 (10 μg/mL) interferenceon the candidate pair (Enzo ALX-804-841-C100-ProSci RF16062(capture-detection)). Serum and plasma samples were incubated with(spiked) and without (unspiked) 150 pg/mL of free LIGHT (in the form ofLIGHT standard recombinant antigen). Said spiked and unspiked sampleswere incubated with DcR3. The % recovery signal was calculated comparedto the control with no interference based on the MFI. The % recoverysignal with the interference is divided by the signal of the control.The signal inhibition value was calculated for unspiked serum 1 (whichhad the most significant reduction). The signal inhibition value wascalculated for unspiked serum 1 (which had the most significantreduction). 78% represents the reduction in signal, which is related(100%-% recovery). That is, Serum l's DcR3 recovery is 22%, representinga 78% reduction in signal, and the anti-LIGHT antibody recovery is 15%,which is 85% reduction in signal. In addition, in the group of LIGHT(150 pg/ml) spiked samples (lower panel), sample serum 1 demonstrates92% inhibition (8% recovery). FIG. 4B: Another spike and recoveryexperiment with the same parameters was performed to assess DcR3interference on a different free LIGHT detecting candidate pair (ProSciRF16062-LSBio LS-C133566-100 (capture-Detection)). FIG. 4C: Graphcharacterizing DcR3 interference with recovery for each candidate pairin native free LIGHT and spiked free LIGHT samples. The relatively low %recovery signal in the native free LIGHT sample for the EnzoALX-804-841-C100-ProSci RF16062 candidate pair indicated that the pairbinds native free LIGHT. In contrast, the relatively little DcR3interference with % recovery signal for the ProSci RF16062-LSBioLS-C133566-100 candidate pair indicates the pair does not as effectivelybind native free LIGHT, even though both candidate pairs bound tonon-native LIGHT standard recombinant antigen to about the same degree.

FIG. 5 shows free LIGHT levels in serum samples from 89 Crohn's Disease(CD) subjects were selected and grouped according to time from illness.89 subjects and 10 healthy controls (gender and age matched) weremeasured using the free-LIGHT assay described herein using the candidateantibody pair. After excluding outliers, 62 samples and 7 controls wereanalyzed. Crohn's Disease subjects showed significantly high serum freeLIGHT levels (527.93 pg/ml, average in subjects of 0-1 month fromillness) than in healthy controls (40.43 pg/ml; P<0.0021). Free LIGHTserum levels also correlated with the disease programs. This suggeststhat Free LIGHT represents a potential target for the treatment of CDand free LIGHT assay can serve as a companion diagnostic for anti-LIGHTtherapy.

FIG. 6 shows serum free LIGHT levels in hospitalized COVID-19 patientsversus healthy controls. Free LIGHT levels in serum were analyzed usingthe Kruskal-Wallis test (non-parametric one-way ANOVA). P-value was<0.0001 indicating higher free LIGHT levels for COVID-19 patients versuscontrols.

FIG. 7 shows serum free LIGHT levels in non-ventilated and intubatedCOVID-19 patients versus healthy controls. Free LIGHT levels in serumwere analyzed using the non-parametric Kruskal-Wallis test. Separatetests were performed for comparison of Non-Ventilated and Intubatedpatients versus Controls. P-values for both tests were <0.0001.

FIG. 8 shows serum free LIGHT levels were compared between healthycontrols over 60 years of age (N=14), subjects over 60 years of age thateventually recovered (N=5), and subjects over 60 years of age thateventually died (N=23) using the Kruskal-Wallis test.

FIG. 9 shows serum free LIGHT levels were compared in subjects in thestudy described in Examples 2 and 3. Square boxes are subjects treatedwith placebo (n=34), circles are subjects treated with the anti-LIGHTmonoclonal antibody(n=36). Mean free LIGHT levels were comparable atbaseline across cohorts. Mean free LIGHT levels reduced dramatically byday 1 after treatment with the anti-LIGHT antibody, but increased in theplacebo treated group. Mean free LIGHT levels were about 100 pg/mLhigher in the patients ≥60 years-old. The pharmacodynamic effect was ontop of standard of care where approximately 90% of patients receivedsystemic corticosteroids.

FIG. 10 shows primary endpoint of the study described in Examples 2 and3: percentage of patients alive and free of respiratory failure at day28 in the anti-LIGHT monoclonal antibody-treated group compared to theplacebo-treated group.

FIG. 11 shows a box plot generated from data from LIGHT testing usingthe free LIGHT assay described herein performed on samples from ARDSpatients and compared to healthy donor LIGHT levels. There are 79healthy control data points.

DETAILED DESCRIPTION OF THE INVENTION

The following definitions are provided to facilitate an understanding ofthe invention. They are not intended to limit the invention in any way.

Definitions

For purposes of the present invention, “a” or “an” entity refers to oneor more of that entity; for example, “a cDNA” refers to one or more cDNAor at least one cDNA. As such, the terms “a” or “an,” “one or more” and“at least one” can be used interchangeably herein. It is also noted thatthe terms “comprising,” “including,” and “having” can be usedinterchangeably. Furthermore, a compound “selected from the groupconsisting of” refers to one or more of the compounds in the list thatfollows, including mixtures (i.e. combinations) of two or more of thecompounds. According to the present invention, an “isolated,” or“biologically pure” molecule is a compound that has been removed fromits natural milieu. As such, the terms “isolated” and “biologicallypure” do not necessarily reflect the extent to which the compound hasbeen purified. An isolated compound of the present invention can beobtained from its natural source, can be produced using laboratorysynthetic techniques or can be produced by any such chemical syntheticroute.

A “coronavirus,” “corona respiratory virus,” or “CoV” are usedinterchangeably herein to refer to a virus belonging to the familyCoronaviridae. Coronaviruses are enveloped, positive-sense RNA virusesof approximately 31 Kb, making these viruses the largest known RNAviruses. Coronaviruses infect a variety of host species, includinghumans and several other vertebrates. These viruses predominantly causerespiratory and intestinal tract infections and induce a wide range ofclinical manifestations. In general, coronaviruses can be classifiedinto low pathogenic CoVs (including human CoVs (hCoVs)) and highlypathogenic CoVs, such as severe acute respiratory syndrome CoV(SARS-CoV) and Middle East respiratory syndrome CoV (MERS-CoV). Lowpathogenic hCoV infect upper airways and cause seasonal mild to moderatecold-like respiratory illnesses in healthy individuals. In contrast, thehighly pathogenic hCoVs (pathogenic hCoV) infect the lower respiratorytract and cause severe pneumonia, which sometimes leads to fatal acutelung injury (ALI) and acute respiratory distress syndrome (ARDS),resulting in high morbidity and mortality. SARS-CoV2 is a type ofcoronavirus. A coronavirus infection as used herein includes any of theabove, if associated with coronavirus. “COVID-19 infection” used hereinmay also refer to a condition or disease caused by SARS-CoV2.

An “acute lung injury” or “ALI” herein refers to an acute lung diseasewith bilateral pulmonary infiltrate in a chest radiograph consistentwith the presence of edema and no clinical evidence of left atrialhypertension; or (if measured) a pulmonary wedge pressure of 18 mmHg orless. Additionally, the ratio of arterial oxygen to the fraction ofinspired oxygen (PaO2/FiO2) must be 300 mmHg or less, regardless of thelevel of positive end-expiratory pressure (PEEP).

“Acute respiratory distress syndrome” or “ARDS” herein refers to themost severe form of ALI, defined by a ratio of arterial oxygen tofraction of inspired oxygen of 200 mmHg or less. The term ARDS is ofteninformally used interchangeably with ALI, but by strict criteria, ARDSshould be reserved for the most severe form of the disease.

“LIGHT” or “TNFSF 14” herein refers to a specific member protein of thetumor necrosis factor superfamily that is expressed by activated Tcells, monocytes-macrophages and additional types of antigen presentingcells. “LIGHT” is an acronym for “homologous to Lymphotoxin, exhibitsInducible expression and competes with HSV Glycoprotein D for binding toHVEM (herpesvirus entry mediator), a receptor expressed on Tlymphocytes.”

“Free LIGHT” or “free (active) LIGHT” herein refers to non-bound formLIGHT (e.g., LIGHT bound to DcR3), which is the active form of LIGHT. Inhumans, free LIGHT is neutralized (inactivated) by DcR3, a uniquesoluble member of the TNFR superfamily, which binds LIGHT in highaffinity and inhibits its interactions with two TNF receptors, HVEM andLTβR. “Bound LIGHT,” or the like, refers to LIGHT that is bound to anatural ligand, optionally wherein the natural ligand is HVEM, LTβR, orDcR3. “Total LIGHT,” or the like, refers to the total amount of freeLIGHT and bound LIGHT.

“Elevated free LIGHT” as used herein refers to a level of free LIGHTdetected in a subject that is higher than a normal control. The normalcontrol can be determined by those of skill in the art as applicable tothe particular situation. In some instances, the normal control is anindustry standard agreed upon by those of skill as being a level orrange of levels that is typical of an individual without aLIGHT-associated condition. In some instances, the normal control is areference level of LIGHT from the same individual taken at a time point,and whether the subject has elevated LIGHT is determined based on asample from that same individual taken at a different, typically later,time point.

The term “antibody” herein is used in the broadest sense and encompassesvarious antibody structures, including but not limited to monoclonalantibodies, polyclonal antibodies, multispecific antibodies (e.g.,bispecific antibodies), and antibody fragments so long as they exhibitthe desired antigen-binding activity. As used herein, the term refers toa molecule comprising at least complementarity-determining region (CDR)1, CDR2, and CDR3 of a heavy chain and at least CDR1, CDR2, and CDR3 ofa light chain, wherein the molecule is capable of binding to antigen.The term antibody includes, but is not limited to, fragments that arecapable of binding antigen, such as Fv, single-chain Fv (scFv), Fab,Fab′, and (Fab′)₂. The term antibody also includes, but is not limitedto, chimeric antibodies, humanized antibodies, human antibodies, andantibodies of various species such as mouse, cynomolgus monkey, etc.

The term “heavy chain” refers to a polypeptide comprising at least aheavy chain variable region, with or without a leader sequence. In someembodiments, a heavy chain comprises at least a portion of a heavy chainconstant region. The term “full-length heavy chain” refers to apolypeptide comprising a heavy chain variable region and a heavy chainconstant region, with or without a leader sequence.

The term “heavy chain variable region” refers to a region comprising aheavy chain complementary determining region (CDR) 1, framework region(FR) 2, CDR2, FR3, and CDR3 of the heavy chain. In some embodiments, aheavy chain variable region also comprises at least a portion of an FR1and/or at least a portion of an FR4. In some embodiments, a heavy chainCDR1 corresponds to Kabat residues 31 to 35; a heavy chain CDR2corresponds to Kabat residues 50 to 65; and a heavy chain CDR3corresponds to Kabat residues 95 to 102. See, e.g., Kabat Sequences ofProteins of Immunological Interest (1987 and 1991, NIH, Bethesda, Md.).

The term “light chain” refers to a polypeptide comprising at least alight chain variable region, with or without a leader sequence. In someembodiments, a light chain comprises at least a portion of a light chainconstant region. The term “full-length light chain” refers to apolypeptide comprising a light chain variable region and a light chainconstant region, with or without a leader sequence. The term “lightchain variable region” refers to a region comprising a light chain CDR1,FR2, HVR2, FR3, and HVR3. In some embodiments, a light chain variableregion also comprises an FR1 and/or an FR4. In some embodiments, a lightchain CDR1 corresponds to Kabat residues 24 to 34; a light chain CDR2corresponds to Kabat residues 50 to 56; and a light chain CDR3corresponds to Kabat residues 89 to 97. See, e.g., Kabat Sequences ofProteins of Immunological Interest (1987 and 1991, NIH, Bethesda, Md.).

A “chimeric antibody” refers to an antibody in which a portion of theheavy and/or light chain is derived from a particular source or species,while the remainder of the heavy and/or light chain is derived from adifferent source or species. In some embodiments, a chimeric antibodyrefers to an antibody comprising at least one variable region from afirst species (such as mouse, rat, cynomolgus monkey, etc.) and at leastone constant region from a second species (such as human, cynomolgusmonkey, etc.). In some embodiments, a chimeric antibody comprises atleast one mouse variable region and at least one human constant region.In some embodiments, a chimeric antibody comprises at least onecynomolgus variable region and at least one human constant region. Insome embodiments, all of the variable regions of a chimeric antibody arefrom a first species and all of the constant regions of the chimericantibody are from a second species.

A “humanized antibody” refers to an antibody in which at least one aminoacid in a framework region of a non-human variable region has beenreplaced with the corresponding amino acid from a human variable region.In some embodiments, a humanized antibody comprises at least one humanconstant region or fragment thereof. In some embodiments, a humanizedantibody is an Fab, an scFv, a (Fab′)₂, etc.

A “human antibody” as used herein refers to antibodies produced inhumans, antibodies produced in non-human animals that comprise humanimmunoglobulin genes, such as XenoMouse®, and antibodies selected usingin vitro methods, such as phage display, wherein the antibody repertoireis based on a human immunoglobulin sequences.

The term “leader sequence” refers to a sequence of amino acid residueslocated at the N terminus of a polypeptide that facilitates secretion ofa polypeptide from a mammalian cell. A leader sequence may be cleavedupon export of the polypeptide from the mammalian cell, forming a matureprotein. Leader sequences may be natural or synthetic, and they may beheterologous or homologous to the protein to which they are attached.

“Percent (%) amino acid sequence identity” and “homology” with respectto a peptide, polypeptide or antibody sequence are defined as thepercentage of amino acid residues in a candidate sequence that areidentical with the amino acid residues in the specific peptide orpolypeptide sequence, after aligning the sequences and introducing gaps,if necessary, to achieve the maximum percent sequence identity, and notconsidering any conservative substitutions as part of the sequenceidentity. Alignment for purposes of determining percent amino acidsequence identity can be achieved in various ways that are within theskill in the art, for instance, using publicly available computersoftware such as BLAST, BLAST-2, ALIGN or MEGALIGN™ (DNASTAR) software.Those skilled in the art can determine appropriate parameters formeasuring alignment, including any algorithms needed to achieve maximalalignment over the full length of the sequences being compared.

The terms “inhibition” or “inhibit” refer to a decrease or cessation ofany event (such as protein ligand binding) or to a decrease or cessationof any phenotypic characteristic or to the decrease or cessation in theincidence, degree, or likelihood of that characteristic. To “reduce” or“inhibit” is to decrease, reduce or arrest an activity, function, and/oramount as compared to a reference. It is not necessary that theinhibition or reduction be complete. For example, in certainembodiments, by “reduce” or “inhibit” is meant the ability to cause anoverall decrease of 20% or greater. In another embodiment, by “reduce”or “inhibit” is meant the ability to cause an overall decrease of 50% orgreater. In yet another embodiment, by “reduce” or “inhibit” is meantthe ability to cause an overall decrease of 75%, 85%, 90%, 95%, orgreater.

“Sample” or “subject sample” or “biological sample” generally refers toa sample which may be tested for a particular molecule. Samples mayinclude but are not limited to cells, body fluids, including blood,serum, plasma, urine, saliva, stool, tears, pleural fluid and the like.

The terms “agent” and “test compound” are used interchangeably hereinand denote a chemical compound, a mixture of chemical compounds, abiological macromolecule, or an extract made from biological materialssuch as bacteria, plants, fungi, or animal (particularly mammalian)cells or tissues. Biological macromolecules include siRNA, shRNA,antisense oligonucleotides, peptides, peptide/DNA complexes, and anynucleic acid based molecule which exhibits the capacity to modulate theactivity of the SNP containing nucleic acids described herein or theirencoded proteins. Agents are evaluated for potential biological activityby inclusion in screening assays described hereinbelow.

A “subject” can be mammalian. In any of the embodiments involving asubject, the subject can be human. In any of the embodiments involving asubject, the subject can be a cow, pig, monkey, sheep, dog, cat, fish,or poultry.

A “pediatric” subject herein is a human of less than 18 years of age,whereas an “adult” subject is 18 years or older.

“Treatment” or “treat” refers to both therapeutic treatment andprophylactic or preventative measures. Those in need of treatmentinclude those already with the disorder as well as those prone to havethe disorder or those in which the disorder is to be prevented. Forpurposes of this invention, beneficial or desired clinical resultsinclude, but are not limited to, alleviation of symptoms, diminishmentof extent of disease, stabilized (i.e., not worsening) state of disease,delay or slowing of disease progression, amelioration or palliation ofthe disease state, and remission (whether partial or total), whetherdetectable or undetectable. “Treatment” can also mean prolongingsurvival as compared to expected survival if not receiving treatment.Those in need of treatment include those already with the condition ordisorder as well as those prone to have the condition or disorder orthose in which the condition or disorder is to be prevented.

The term “effective amount” or “therapeutically effective amount” refersto an amount of a drug effective for treatment of a disease or disorderin a subject, such as to partially or fully relieve one or moresymptoms. In some embodiments, an effective amount refers to an amounteffective, at dosages and for periods of time necessary, to achieve thedesired therapeutic or prophylactic result.

Identification of Biomarker LIGHT

LIGHT (TNFSF14) is an important regulatory cytokine, which serves ascritical factor in orchestrating a cytokine storm and pulmonary failureassociated with pathogen-mediated infection, including viral andbacterial infections including coronavirus (e.g., COVID-19).

In some embodiments, methods for detecting free (active) LIGHT in ahuman subject are provided. In some embodiments, the subject'sbiological sample (e.g., serum) is analyzed for free LIGHT. In someembodiments, the results provide a basis for understanding whether ananti-LIGHT therapy may be provided and will be effective. For instance,if an elevated level of free LIGHT (e.g., a level of free LIGHT abovewhat is expected, for example, using a normal control) then the subjectmay be diagnosed with a condition associated with elevated free LIGHTand/or be deemed to be a suitable candidate for treatment with ananti-LIGHT antibody. In some embodiments, the anti-LIGHT antibody is anantibody neutralizing LIGHT. In some embodiments, methods for detectingfree (active) LIGHT in a subject's biological sample (e.g., serum) areprovided, wherein the results provide a basis for understanding whetheran anti-LIGHT therapy may be provided and will be effective. In someembodiments, detection of free LIGHT above a normal control indicatesthat an anti-LIGHT therapy may be provided and effective.

In some embodiments, the condition associated with elevated free LIGHTwith which the subject may be diagnosed is Crohn's Disease or aninflammatory condition associated with Crohn's Disease. In someembodiments, the condition associated with elevated free LIGHT withwhich the subject may be diagnosed is a coronavirus infection. In someembodiments, the coronavirus infection is a moderate or a severecoronavirus infection. In some embodiments, conditions associated withelevated free LIGHT include any one or more of inflammation, optionallywherein the inflammation is hyperinflammation, immune dysregulation thatleads to multisystem organ failure, acute lung injury (ALI), optionallywherein the ALI is associated with a bacterial or viral infection,including coronavirus infection, acute respiratory distress syndrome(ARDS), optionally wherein the ARDS is associated with a bacterial orviral infection, including coronavirus infection, cytokine storm thatdrives tissue injury and vascular permeability, post-infection pulmonaryfibrosis, and pneumonia, optionally wherein the pneumonia is associatedwith a bacterial or viral infection, including coronavirus infection. Insome embodiments, the condition associated with elevated free LIGHT ismild, moderate, or severe coronavirus infection, optionally wherein thecoronavirus infection is a COVID-19 infection. In some embodiments, theCOVID-19 infection is associated with ALI or ARDS in the subject. Insome embodiments, the COVID-19 infection is associated with cytokinestorm that drives tissue injury and vascular permeability in the lungsand post-infection pulmonary fibrosis. In some embodiments, thecoronavirus infection is MERS-CoV, SARS-CoV, or SARS-CoV2/COVID-19.

In some embodiments, methods for diagnosing a condition associated withelevated free LIGHT in a subject are provided, wherein the level of free(active) LIGHT in a biological sample is detected. If the levels areabove a normal control, then the subject is diagnosed as having acondition associated with elevated free LIGHT. In some embodiments, thecondition associated with elevated free LIGHT to be detected is Crohn'sDisease or an inflammatory condition associated with Crohn's Disease. Insome embodiments, the condition associated with elevated free LIGHT tobe detected is a virus infection. In some embodiments, the conditionassociated with elevated free LIGHT is a coronavirus infection. In someembodiments, the condition associated with elevated free LIGHT to bedetected is a moderate or a severe coronavirus infection. In someembodiments, the condition associated with elevated free LIGHT to bedetected is a mild, moderate, or severe COVID-19 infection. In someembodiments, the COVID-19 infection is associated with ALI or ARDS inthe subject. In some embodiments, the coronavirus infection is MERS-CoVor SARS-CoV.

In some embodiments, methods for detecting free LIGHT in a biologicalsample from a subject may be conducted with a biological samplecomprising blood, urine, serum, plasma, feces, or gastric lavage bodilyfluid samples, or cell samples such as white blood cells or mononuclearcells.

In some embodiments, the method of detecting free (active) LIGHT insubject is performed by:

-   -   (a) contacting the biological sample with at least one        anti-LIGHT antibody;    -   (b) incubating the biological sample to allow the anti-LIGHT        antibody to bind to free LIGHT; and    -   (c) determining the presence of complexes formed between the        anti-LIGHT antibody and free LIGHT in the biological sample.

This method can further comprise the step of diagnosing the subject ashaving a condition associated with elevated free LIGHT and/oradministering an anti-LIGHT antibody.

Various methods known in the art for detecting specific antibody-antigenbinding can be used. Exemplary immunoassays which can be conductedinclude fluorescence polarization immunoassay (FPIA), fluorescenceimmunoassay (FIA), enzyme immunoassay (EIA), nephelometric inhibitionimmunoassay (NIA), enzyme linked immunosorbent assay (ELISA), andradioimmunoassay (MA), competition assay, and sandwich method.

An indicator moiety, or label group, can be attached to the subjectantibodies and is selected to meet the needs of various uses of themethod which are often dictated by the availability of assay equipmentand compatible immunoassay procedures. Appropriate labels include,without limitation, radionuclides (for example 125I, 131I, 35S, 3H, or32P), enzymes (for example, alkaline phosphatase, horseradishperoxidase, luciferase, or β-galactosidase), fluorescent moieties orproteins (for example, fluorescein, rhodamine, phycoerythrin, GFP, orBFP), or luminescent moieties (for example, Qdot™ nanoparticles suppliedby the Quantum Dot Corporation, Palo Alto, Calif.).

General techniques to be used in performing the various immunoassaysnoted above are known to those of ordinary skill in the art.

ELISA assays are generally known to the skilled artisan and can bedesigned to determine serum LIGHT levels. In one exemplary embodiment,blood is collected, and the serum is isolated. If no kit is available,an ELISA can be developed using plates that are pre-coated with captureantibody specific for the LIGHT one is measuring. The plate is nextincubated at room temperature for a period of time before washing.Enzyme-anti-LIGHT antibody conjugate is added and incubated. Unboundantibody conjugate is removed, and the plate washed before the additionof the chromogenic substrate solution that reacts with the enzyme. Theplate is read on an appropriate plate reader at an absorbance specificfor the enzyme and substrate used.

The competition method compares the competitive binding of an antigen ina sample and a known amount of a labeled antigen to the monoclonalantibody of the present invention. To carry out an immunological assaybased on the competition method, a sample containing an unknown amountof the target antigen is added to a solid substrate to which themonoclonal antibody of the present invention is coated physically orchemically by known means, and the reaction is allowed to proceed.Simultaneously, a predetermined amount of the pre-labeled target antigenis added and the reaction is allowed to proceed. After incubation, thesolid substrate is washed and the activity of the labeling agent boundto the solid substrate is measured.

In the sandwich method, the target antigen in a sample is sandwichedbetween the immobilized monoclonal antibody and the labeled monoclonalantibody, then a labeling substrate such as an enzyme is added,substrate color changes are detected, and thereby detecting the presenceof the antigen. To carry out an immunological assay based on thesandwich method, for instance, a sample containing an unknown amount ofthe target antigen is added to a solid substrate to which the monoclonalantibody of the present invention is coated physically or chemically byknown means, and the reaction is allowed to proceed. Thereafter, thelabeled monoclonal antibody of the invention is added and the reactionis allowed to proceed. After incubation, the solid substrate is washedand the activity of the labeling agent bound to the solid substrate ismeasured.

In some embodiments, a first antibody is used for a diagnostic and asecond antibody is used as a therapeutic. In some embodiments, the firstand second antibodies are different. In some embodiments, the first andsecond antibodies can both bind to the antigen at the same time, bybinding to separate epitopes.

Methods of Treating with Anti-LIGHT Antibody

The present invention provides a method of treating subjects havingelevated free LIGHT, including subjects having a condition associatedwith elevated free LIGHT with an anti-LIGHT antibody. In someembodiments, the anti-LIGHT antibody is an antibody neutralizing LIGHT.In some embodiments, the condition associated with elevated free LIGHTis a coronavirus infection, including COVID-19. In some embodiments, thecondition associated with elevated free LIGHT is Crohn's Disease or aninflammatory condition associated with Crohn's Disease. The treatmentcan be done with or without a diagnostic test for detecting elevatedfree LIGHT in a subject's sample.

In some embodiments, a biological sample from the subject is firsttested for the presence and level of free LIGHT in a threshold stepbefore the subject is treated. In such embodiments, the method oftreating comprises contacting a biological sample from a subject with atleast one first anti-LIGHT antibody, incubating the biological sample toallow the anti-LIGHT antibody to bind to free LIGHT, determining thepresence of complexes formed between the anti-LIGHT antibody and freeLIGHT in the biological sample, and finally, based on the positiveresults of the detecting step and a finding of elevated free LIGHT,administering to the subject an effective amount of a second anti-LIGHTantibody, wherein the first and the second antibody differ.

In some embodiments, the subject is treated without a threshold testingstep. In such cases, it may have been predetermined that the subjectcould benefit from an anti-LIGHT therapy or the anti-LIGHT antibody isbeing administered for prevention. In such embodiments, the method oftreating comprises administering to a subject having a conditionassociated with elevated free LIGHT an effective amount of an anti-LIGHTantibody.

In some embodiments of the method of treating, the condition associatedwith elevated free LIGHT is Crohn's Disease or an inflammatory conditionassociated with Crohn's Disease.

In some embodiments of the method of treating, the condition associatedwith elevated free LIGHT is a coronavirus infection. In someembodiments, the coronavirus infection is a COVID-19 infection. In someembodiments, the subject has a respiratory disease, optionally caused bya virus, bacteria or fungus. In some embodiments, the subject has arespiratory disease caused by coronavirus infection. In someembodiments, the subject has a respiratory disease caused by COVID-19.In some embodiments, the subject has pneumonia. In some embodiments, thesubject has acute lung injury (ALI). In some embodiments, the subjecthas acute respiratory distress syndrome (ARDS). In some embodiments,pneumonia is associated with coronavirus infection. In some embodiments,ALI or ARDS is associated with coronavirus infection. In someembodiments, pneumonia is associated with COVID-19. In some embodiments,the subject has acute lung injury (ALI) associated with COVID-19. Insome embodiments, the subject has acute respiratory distress syndrome(ARDS) associated with COVID-19. In some embodiments, the subject has amild coronavirus infection. In some embodiments, the subject has amoderate coronavirus infection. In some embodiments, the subject has asevere coronavirus infection. In some embodiments, a method of treatingsevere COVID-19 pneumonia comprising administering an anti-LIGHTantibody to a subject in need thereof is provided. In some embodiments,a method of treating acute inflammatory disease associated with COVID-19pneumonia comprising administering an anti-LIGHT antibody to a subjectin need thereof is provided. In some embodiments, a method of treatingrespiratory failure associated with COVID-19 pneumonia comprisingadministering an anti-LIGHT antibody to a subject in need thereof isprovided. In some embodiments, a method of treating cytokine stormcomprising administering an anti-LIGHT antibody to a subject in needthereof is provided. In some embodiments, a method of treating adysregulated hyperimmune response (sometimes referred to as “cytokinestorm”) comprising administering an anti-LIGHT antibody to a subject inneed thereof is provided. In some embodiments, a method of treatingAcute Respiratory Disease Syndrome (ARDS) comprising administering ananti-LIGHT antibody to a subject in need thereof is provided.

In some embodiments, the administration of the anti-LIGHT antibodysuppresses T cell activation. In some embodiments, the administration ofthe anti-LIGHT antibody suppresses increased expression of cytokines(e.g., a cytokine storm). In some embodiments, the administration of theanti-LIGHT antibody suppresses increased expression of cytokines (e.g.,a cytokine storm) that is caused by a viral and/or bacterial infection,e.g., from a coronavirus infection. In some embodiments theadministration of the anti-LIGHT antibody prevents or treats cytokinestorm caused by a virus and/or bacterial infection, such as by acoronavirus infection. In some embodiments, the cytokine storm can drivetissue injury and vascular permeability in the lungs. Cytokine storm canlead to ARDS and ALI associated with coronavirus (e.g., COVID-19)infection. In some embodiments, the administration of the anti-LIGHTantibody prevents or treats post-infection pulmonary fibrosis,optionally associated with coronavirus infection. In some embodiments,the administration of the anti-LIGHT antibody reduces the subject's riskof mortality or morbidity. In some embodiments, the administration ofthe anti-LIGHT antibody prevents progression to ARDS in a subject withpneumonia associated with a coronavirus infection (e.g., COVID-19infection). In some embodiments, the administration of the anti-LIGHTantibody prevents progression of ALI associated with a coronavirusinfection, including (e.g., COVID-19 infection) to ARDS. In someembodiments, the administration of the anti-LIGHT antibody prevents ortreats ARDS. In some embodiments, the administration of the anti-LIGHTantibody prevents the need for ventilation/intubation of the subject.

The methods of treating in the present invention may be carried outthrough conventional administration routes, including withoutlimitation, the oral, buccal, sublingual, ocular, topical, parenteral,rectal, intracisternal, intravaginal, intraperitoneal, intravesical,local (e.g., powder, ointment, or drop), or nasal routes. The term“parenteral” includes subcutaneous, intravenous, intramuscular, orinfusion. In certain embodiments, it may be appropriate to administerthe agent in a continuous infusion or as a subcutaneous injection everyday, every two or several days, or once a week, or every several weeks,or once a month, or once every several months, or at a time intervalwithin a range defined by any two of the aforementioned intervals.

In some embodiments, the anti-LIGHT antibody is administered to asubject already receiving another therapy. In the case of a subject withCOVID-19, the other therapy may be any therapy approved or being testedto treat or ameliorate a symptom of COVID-19. Such therapies include,but are not limited to, remdesivir, corticosteroids, andhydroxychloroquine. In some embodiments, the other therapy continues (onits normal course) during treatment with the anti-LIGHT antibody. Insome embodiments, the other therapy is discontinued during treatmentwith the anti-LIGHT antibody. In some embodiments, the subject hasCOVID-19 and is receiving a high dose of corticosteroid and ananti-LIGHT antibody. In some embodiments, subject receiving thecombination high dose corticosteroid and anti-LIGHT antibody have betteroutcomes than those not receiving the combination.

In some embodiments, the dose of anti-LIGHT antibody is about 16 mg/kgwith a maximum dose of 1,200 mg. In some embodiments, the dose is asingle administration. In some embodiments, the dose is more than asingle administration.

In some embodiments, administration of the anti-LIGHT antibody reducesserum free-LIGHT levels in the subject by 85% or more, for example,compared to free-LIGHT levels in the subject prior to administration ofthe anti-LIGHT antibody or compared to subjects who are not administeredthe anti-LIGHT antibody. In some embodiments, the reduction in serumfree-LIGHT levels in the subject by 85% or more occurs in less than 5days or less than 1 day after administration of the anti-LIGHT antibody.In some embodiments, the anti-LIGHT antibody administered to the subjectreduces the subject's risk of mortality by equal to or greater than 50%at 60 days after administration. In some embodiments, the anti-LIGHTantibody administered to the subject reduces the subject's risk ofmortality by equal to or greater than 50% at 28 days afteradministration. In some embodiments, the subject is 60 years of age orolder. In some embodiments, the anti-LIGHT antibody administered to thesubject shortens the length of the subject's hospital stay compared tosubjects receiving standard of care treatment. In some embodiments, theanti-LIGHT antibody administered to the subject reduces the subject'srisk of respiratory failure.

In some embodiments, the anti-LIGHT antibody is administered as a doseof from about 1.0 mg/kg to about 3.0 mg/kg. In some embodiments, theanti-LIGHT antibody is administered as a dose of about 1.0, about 1.1,about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about1.8, about 1.9, about 2.0, about 2.1, about 2.2, about 2.3, about 2.4,about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, or about 3.0mg/kg. In some embodiments, the anti-LIGHT antibody is administeredevery day, every 2 days, every 3 days, every 4 days, every 5 days, every6 days, every week, every two weeks, every three weeks, or every fourweeks. In some embodiments, the anti-LIGHT antibody is administered as adose of from about 1.0 mg/kg to about 3.0 mg/kg every 14 days. In someembodiments, the anti-LIGHT antibody is administered as a dose of 1.0mg/kg every 14 days. In some embodiment, the anti-LIGHT antibody isadministered as a dose of 3.0 mg/kg every 14 days.

In some embodiments, the anti-LIGHT antibody is administered to asubject who has failed treatment with an approved therapeutic dose of ananti-TNFα monoclonal antibody treatment with either no initial responseor an initial response to induction with subsequent lost response.

Treatment of Crohn's Disease

The primary clinical target for CD treatment is the resolution ofsymptoms. (Shi and Ng, The state of the art on treatment of Crohn'sdisease, J. Gastroenterol., 2018:53; 989-998). The standard index forassessing symptoms is the Crohn's Disease Activity Index (CDAI), whichincludes eight variables: the number of liquid stools, the extent ofabdominal pain, general well-being, the occurrence of extraintestinalsymptoms, the need for antidiarrhea drugs, the presence of abdominalmasses, hematocrit, and body weight. (Shi and Ng 2018). Control ofsymptoms does not appear to significantly alter the natural course ofthe disease. (Shi and Ng 2018).

Mucosal healing is becoming another target in clinical practice. TheSimple Endoscopic Score for Crohn's Disease (SES-CD) is one of the mostcommon endoscopic scoring systems for CD. The SES-CD is scored based onfour endoscopic variables (presence and size of ulcers, extent ofulcerated surface, extent of affected surface, and presence and type ofnarrowing) in the same five segments. An SES-CD score below 2 isregarded as endoscopic remission. A decrease from baseline of at least50% in an SES-CD score has been used as the definition for endoscopicresponse. There is currently no set, universal definition for mucosalhealing, but the absence of ulceration at ileocolonoscopy has beenadopted as the endoscopic endpoint for CD. (Shi and Ng 2018).

A Patient reported outcome is a measurement derived directly from apatient about any aspect of their health status and has the potential tobecome a treatment endpoint for CD. Several scales have been used toassess patients' perspectives towards the disease, including theInflammatory Bowel Disease Questionnaire (IBD-Q). (Shi and Ng 2018).

In some embodiments, administration of the anti-LIGHT antibody reducesthe subject's CDAI score, for example, compared to the subject's CDAIscore prior to administration of the anti-LIGHT antibody or compared tosubjects who are not administered the anti-LIGHT antibody. In someembodiments, administration of the anti-LIGHT antibody decreases thesubject's SES-CD score, for example, compared to the subject's SES-CDscore prior to administration of the anti-LIGHT antibody or compared tosubjects who are not administered the anti-LIGHT antibody. In someembodiments, administration of the anti-LIGHT antibody results in thesubject's SES-CD score of below 2. In some embodiments, administrationof the anti-LIGHT antibody increases the subject's IBD-Q score, forexample, compared to the subject's IBD-Q score prior to administrationof the anti-LIGHT antibody or compared to subjects who are notadministered the anti-LIGHT antibody. In some embodiments,administration of the anti-LIGHT antibody results in the subject's IBD-Qscore of 170 or higher. In some embodiments, administration of theanti-LIGHT antibody results in an increase in the subject's IBD-Q scoreof at least 16 points. In some embodiments, administration of theanti-LIGHT antibody results in an increase in the subject's IBD-Q scoreof at least 32 points.

Stages of COVID-19 Disease Progression

The stages of COVID-19 Disease Progression are shown in FIG. 1 ofSiddiqi, H. K., et al. COVID-19 illness in native and immunosuppressedstates: A clinical-therapeutic staging proposal. J Heart Lung Transplant39(5), 405-407 (2020). The disease can broadly be divided into threestages: Early Infection, Pulmonary Phase, and Hyperinflammation Phase.Stage I is the viral response phase, Stage III is the host inflammatoryresponse phase, and Stage II is a combination of the two. As patientsprogress from Early Infection (Stage I) to the Pulmonary Phase (StageII) at which pulmonary symptoms appear, the host inflammatory responseemerges. It is at this point where, in some patients, dysregulation ofthe inflammatory response occurs. This dysregulation of the inflammatoryresponse may be accompanied by elevated free LIGHT levels in biologicalfluids, such as serum. This stage, the Pulmonary Phase, is the optimalstage at which to administer an anti-LIGHT antibody to correct thisdysregulation and avert cytokine storm and progression to ARDS, whichoccurs in the Hyperinflammation Phase (Stage III).

Patients in Stage I of COVID-19 are considered to have mild disease.Patients in Stage II of COVID-19 are considered to have mild to moderatedisease. Patients in Stage III of COVID-19 are considered to have severedisease.

In some embodiments, the anti-LIGHT antibody is administered to asubject with mild coronavirus (e.g., COVID-19). In some embodiments, theanti-LIGHT antibody is administered during the early infection stage ofa coronavirus. In some embodiments, the anti-LIGHT antibody isadministered during Stage I of a coronavirus. In some embodiments, theanti-LIGHT antibody is administered while the subject in need thereofhas mild constitutional symptoms, a fever >99.6° F., dry cough,diarrhea, or headache. In some embodiments, the subject in need thereofhas lymphopenia, increased prothrombin time, increased D-dimer and LDH(mild). In some embodiments, the administration of the anti-LIGHTantibody treats Stage I of a coronavirus. In some embodiments, theadministration of the anti-LIGHT antibody prevents progression of thecoronavirus to Stage II. Staging refers to FIG. 1 of Siddiqi, H. K., etal. (2020), incorporated herein in its entirety.

In some embodiments, the anti-LIGHT antibody of the method isadministered to a subject with moderate coronavirus (e.g., COVID-19). Insome embodiments, the anti-LIGHT antibody is administered during thePulmonary Phase of a coronavirus. In some embodiments, the anti-LIGHTantibody is administered during Stage II of a coronavirus. In someembodiments of the invention, the anti-LIGHT antibody is administeredduring Stage IIA or Stage IIB of a coronavirus. In some embodiments, theanti-LIGHT antibody is administered while the subject in need thereof isexhibiting shortness of breath or hypoxia. In some embodiments, theanti-LIGHT antibody is administered while the subject in need thereofmeets the clinical criteria for ALI. In some embodiments, the subject inneed thereof has abnormal chest imaging, transaminitis, or low-normalprocalcitonin. In some embodiments, the patient is notventilated/intubated. In some embodiments, the administration of theanti-LIGHT antibody treats Stage II of a coronavirus. In someembodiments, the administration of the anti-LIGHT antibody preventsprogression of the coronavirus to Stage III.

In some embodiments, the anti-LIGHT antibody is administered to asubject with severe coronavirus (e.g., COVID-19). In some embodiments,the anti-LIGHT antibody is administered during the HyperinflammationPhase of a coronavirus. In some embodiments, the anti-LIGHT antibody isadministered during Stage III of a coronavirus. In some embodiments, theanti-LIGHT antibody is administered while the subject in need thereofhas ARDS, systemic inflammatory response syndrome (SIRS)/Shock, orCardiac Failure. SIRS is a complex immune response to insultcharacterized by widespread inflammation within the body. Bothinfectious and non-infectious causes of SIRS have been identified. Insome embodiments, the patient is ventilated/intubated. In someembodiments, the administration of the anti-LIGHT antibody treats StageIII of a coronavirus.

Anti-LIGHT Antibodies

In some embodiments, an anti-LIGHT antibody is utilized for bothdetection/diagnostic and therapeutic purposes, as well as in the assaysdescribed herein. The anti-LIGHT antibody used for detection ordiagnostic purposes is different from the antibody used for therapeuticpurposes (even in the same subject).

The anti-LIGHT antibody useful for therapeutic purposes may comprise theCDR sequences of the E1, E13, E63, F19, or F23 antibodies, which areprovided in WO 2008/027338 and U.S. Pat. Nos. 8,058,402 B2, 8,461,307B2, and 8,974,787 B2, each of which is incorporated herein by reference.The anti-LIGHT antibody useful for detection/diagnostic purposes is notthat same as that which is used for therapeutic purposes.

In some embodiments, the anti-LIGHT antibody comprises a heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 2, 3, and4, respectively. In some embodiments, the anti-LIGHT antibody comprisesa light chain comprising three CDR sequences comprising each of SEQ IDNOs: 5, 6, and 7, respectively. In some embodiments, the antibodycomprises a heavy chain and a light chain, the heavy chain comprisingthree CDR sequences comprising each of SEQ ID NOs: 2, 3, and 4,respectively, and the light chain comprising three CDR sequencescomprising each of SEQ ID NOs: 5, 6, and 7, respectively.

In some embodiments, the anti-LIGHT antibody comprises a heavy chainvariable region sequence comprising SEQ ID NO: 8 or that is at least85%, at least 90%, at least 95%, at least 98%, or at least 99% identicalto SEQ ID NO:8. In some embodiments, the anti-LIGHT antibody comprises alight chain variable region sequence comprising SEQ ID NO:9 or that isat least 85%, at least 90%, at least 95%, at least 98%, or at least 99%identical to SEQ ID NO:9. In some embodiments, the anti-LIGHT antibodycomprises a heavy chain comprising SEQ ID NO: 8 or that is at least 85%,at least 90%, at least 95%, at least 98%, or at least 99% identical toSEQ ID NO:8. In some embodiments, the anti-LIGHT antibody comprises alight chain comprising SEQ ID NO:9 or that is at least 85%, at least90%, at least 95%, at least 98%, or at least 99% identical to SEQ IDNO:9. In some embodiments, the anti-LIGHT antibody comprises both aheavy chain comprising SEQ ID NO: 8 or that is at least 85%, at least90%, at least 95%, at least 98%, or at least 99% identical to SEQ IDNO:8 and a light chain comprising SEQ ID NO:9 or that is at least 85%,at least 90%, at least 95%, at least 98%, or at least 99% identical toSEQ ID NO:9.

In some embodiments, the anti-LIGHT antibody comprises a heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 10, 11,and 12, respectively. In some embodiments, the anti-LIGHT antibodycomprises a light chain comprising three CDR sequences comprising eachof SEQ ID NOs: 13, 14, and 15, respectively. In some embodiments, theantibody comprises a heavy chain and a light chain, the heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 10, 11,and 12, respectively, and the light chain comprising three CDR sequencescomprising each of SEQ ID NOs: 13, 14, and 15, respectively.

In some embodiments, the anti-LIGHT antibody comprises a heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 16, 17,and 18, respectively. In some embodiments, the anti-LIGHT antibodycomprises a light chain comprising three CDR sequences comprising eachof SEQ ID NOs: 19, 20, and 21, respectively. In some embodiments, theantibody comprises a heavy chain and a light chain, the heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 16, 17,and 18, respectively, and the light chain comprising three CDR sequencescomprising each of SEQ ID NOs: 19, 20, and 21, respectively.

In some embodiments, the anti-LIGHT antibody comprises a heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 22, 23,and 24, respectively. In some embodiments, the anti-LIGHT antibodycomprises a light chain comprising three CDR sequences comprising eachof SEQ ID NOs: 25, 26, and 27, respectively. In some embodiments, theantibody comprises a heavy chain and a light chain, the heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 22, 23,and 24, respectively, and the light chain comprising three CDR sequencescomprising each of SEQ ID NOs: 25, 26, and 27, respectively.

In some embodiments, the anti-LIGHT antibody comprises a heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 28, 29,and 30, respectively. In some embodiments, the anti-LIGHT antibodycomprises a light chain comprising three CDR sequences comprising eachof SEQ ID NOs: 31, 32, and 33, respectively. In some embodiments, theantibody comprises a heavy chain and a light chain, the heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 28, 29,and 30, respectively, and the light chain comprising three CDR sequencescomprising each of SEQ ID NOs: 31, 32, and 33, respectively.

In some embodiments, the anti-LIGHT antibody comprises a heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 34, 35,and 36, respectively. In some embodiments, the anti-LIGHT antibodycomprises a light chain comprising three CDR sequences comprising eachof SEQ ID NOs: 37, 38, and 39, respectively. In some embodiments, theantibody comprises a heavy chain and a light chain, the heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 34, 35,and 36, respectively, and the light chain comprising three CDR sequencescomprising each of SEQ ID NOs: 37, 38, and 39, respectively.

In some embodiments, the anti-LIGHT antibody comprises a heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 40, 41,and 42, respectively. In some embodiments, the anti-LIGHT antibodycomprises a light chain comprising three CDR sequences comprising eachof SEQ ID NOs: 43, 44, and 45, respectively. In some embodiments, theantibody comprises a heavy chain and a light chain, the heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 40, 41,and 42, respectively, and the light chain comprising three CDR sequencescomprising each of SEQ ID NOs: 43, 44, and 45, respectively.

In some embodiments, the anti-LIGHT antibody comprises a heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 46, 47,and 48, respectively. In some embodiments, the anti-LIGHT antibodycomprises a light chain comprising three CDR sequences comprising eachof SEQ ID NOs: 49, 50, and 51, respectively. In some embodiments, theantibody comprises a heavy chain and a light chain, the heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 46, 47,and 48, respectively, and the light chain comprising three CDR sequencescomprising each of SEQ ID NOs: 49, 50, and 51, respectively.

In some embodiments, the anti-LIGHT antibody may comprise the CDRsequences of the antibodies, which are described in US2013/0323240 andU.S. Pat. No. 8,524,869 B2, which are incorporated herein by reference.For example, in some embodiments, the anti-LIGHT antibody comprises aheavy chain comprising three CDR sequences comprising each of SEQ IDNOs: 52, 53, and 54, respectively. In some embodiments, the anti-LIGHTantibody comprises a light chain comprising three CDR sequencescomprising each of SEQ ID NOs: 55, 56, and 57, respectively. In someembodiments, the antibody comprises a heavy chain and a light chain, theheavy chain comprising three CDR sequences comprising each of SEQ IDNOs: 52, 53, and 54, respectively, and the light chain comprising threeCDR sequences comprising each of SEQ ID NOs: 55, 56, and 57,respectively.

In some embodiments, the anti-LIGHT antibody comprises a heavy chainvariable region sequence comprising SEQ ID NO:58 or that is at least85%, at least 90%, at least 95%, at least 98%, or at least 99% identicalto SEQ ID NO:58. In some embodiments, the anti-LIGHT antibody comprisesa light chain variable region sequence comprising SEQ ID NO:59 or thatis at least 85%, at least 90%, at least 95%, at least 98%, or at least99% identical to SEQ ID NO:59. In some embodiments, the anti-LIGHTantibody comprises a heavy chain comprising SEQ ID NO:58 or that is atleast 85%, at least 90%, at least 95%, at least 98%, or at least 99%identical to SEQ ID NO:58. In some embodiments, the anti-LIGHT antibodycomprises a light chain comprising SEQ ID NO:59 or that is at least 85%,at least 90%, at least 95%, at least 98%, or at least 99% identical toSEQ ID NO:59. In some embodiments, the anti-LIGHT antibody comprisesboth a heavy chain comprising SEQ ID NO:58 or that is at least 85%, atleast 90%, at least 95%, at least 98%, or at least 99% identical to SEQID NO:58 and a light chain comprising SEQ ID NO:59 or that is at least85%, at least 90%, at least 95%, at least 98%, or at least 99% identicalto SEQ ID NO:59.

In some embodiments, the anti-LIGHT antibody may comprise a heavy chainand a light chain together comprising one of the following sets ofCDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 sequences describedin the sequence listing from US2013/0323240: SEQ ID NOs: 18, 19, 20 andSEQ ID NOs: 38, 41, 42 of US2013/0323240; SEQ ID NOs: 18, 19, 21 and SEQID NOs: 39, 41, 42 of US2013/0323240; SEQ ID NOs: 18, 19, 22 and SEQ IDNOs: 40, 41, 42 of US2013/0323240; SEQ ID NOs: 23, 24, 25 and SEQ IDNOs: 43, 44, 45 of US2013/0323240; SEQ ID NOs: 26, 27, 28 and SEQ IDNOs: 46, 47, 48 of US2013/0323240; SEQ ID NOs: 29, 30, 31 and SEQ IDNOs: 49, 50, 51 of US2013/0323240; SEQ ID NOs: 32, 33, 34 and SEQ IDNOs: 52, 53, 54 of US2013/0323240; and SEQ ID NOs: 35, 36, 37 and SEQ IDNOs: 55, 50, 51 of US2013/0323240.

In some embodiments, the anti-LIGHT antibody comprises the CDR sequencesof the 18E04, 98C07, 1C02, 1C06, 13H04, 31A10, 98C07, 42A02, 29C02,14B09, 117C06, 114F05, and 62C01 antibodies described in WO 2015/107331,which is also incorporated by reference herein.

For example, in some embodiments, the anti-LIGHT antibody comprises aheavy chain comprising three CDR sequences comprising each of SEQ IDNOs: 60, 61, and 62, respectively. In some embodiments, the anti-LIGHTantibody comprises a light chain comprising three CDR sequencescomprising each of SEQ ID NOs: 63, 64, and 65, respectively. In someembodiments, the antibody comprises a heavy chain and a light chain, theheavy chain comprising three CDR sequences comprising each of SEQ IDNOs: 60, 61, and 62, respectively, and the light chain comprising threeCDR sequences comprising each of SEQ ID NOs: 63, 64, and 65,respectively.

In some embodiments, the anti-LIGHT antibody comprises a heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 66, 67,and 68, respectively. In some embodiments, the anti-LIGHT antibodycomprises a light chain comprising three CDR sequences comprising eachof SEQ ID NOs: 69, 70, and 71, respectively. In some embodiments, theantibody comprises a heavy chain and a light chain, the heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 66, 67,and 68, respectively, and the light chain comprising three CDR sequencescomprising each of SEQ ID NOs: 69, 70, and 71, respectively.

In some embodiments, the anti-LIGHT antibody comprises a heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 72, 73,and 74, respectively. In some embodiments, the anti-LIGHT antibodycomprises a light chain comprising three CDR sequences comprising eachof SEQ ID NOs: 75, 76, and 77, respectively. In some embodiments, theantibody comprises a heavy chain and a light chain, the heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 72, 73,and 74, respectively, and the light chain comprising three CDR sequencescomprising each of SEQ ID NOs: 75, 76, and 77, respectively.

In some embodiments, the anti-LIGHT antibody comprises a heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 78, 79,and 80, respectively. In some embodiments, the anti-LIGHT antibodycomprises a light chain comprising three CDR sequences comprising eachof SEQ ID NOs: 81, 82, and 83, respectively. In some embodiments, theantibody comprises a heavy chain and a light chain, the heavy chaincomprising three CDR sequences comprising each of SEQ ID NOs: 78, 79,and 80, respectively, and the light chain comprising three CDR sequencescomprising each of SEQ ID NOs: 81, 82, and 83, respectively.

Kits and Articles of Manufacture

Any of the aforementioned methods can be implemented via kits for thedetection of free LIGHT and/or the diagnosis and/or treatment of acondition associated with elevated free LIGHT, including a viral and/orbacterial infection, such as a coronavirus, including COVID-19. The kitmay contain an antibody, one or more non-naturally occurring detectablelabels, marker, or reporter, a pharmaceutically acceptable carrier, aphysiologically acceptable carrier, instructions for use, a container, avessel for administration, an assay substrate, or any combinationthereof.

In some embodiments, the kit is for use in a method of detecting freeLIGHT in a biological sample (as opposed to total or bound LIGHT).

In some embodiments, the kit is for use in a method of detecting freeLIGHT in a biological sample from a subject having or suspected ofhaving coronavirus infection, ALI, ARDS, or pulmonary fibrosis.

In some embodiments, the kit is for use in a method of detecting freeLIGHT in a biological sample from a subject having or suspected ofhaving Crohn's Disease or an inflammatory condition associated withCrohn's Disease.

In some embodiments, the kit is for use in method of detecting elevatedfree LIGHT in a biological sample from a subject, optionally wherein thesubject is suspected of having a coronavirus infection and also fortreating the subject after diagnosis by administering an anti-LIGHTantibody in an effective amount.

In some embodiments, the kit for use in one of the above methods issuitable for use in a subject with mild, moderate, or severe Crohn'sDisease or a mild, moderate, or severe inflammatory condition associatedwith Crohn's Disease.

In some embodiments, the kit for use in one of the above methods issuitable for a subject with a mild, moderate, or severe coronavirusinfection. In some embodiments, the mild, moderate, or severecoronavirus infection is a COVID-19 infection. In some embodiments, thecoronavirus (including COVID-19) infection is associated withrespiratory disease. In some embodiments, the coronavirus (includingCOVID-19) infection is associated with ALI or ARDS.

In some embodiments, the kit for use in a method of detecting and/ortreating comprises a solid phase to which the anti-LIGHT antibodyreagent is attached. In some embodiments, the kit for use in a method ofdetecting and/or treating comprises a solid phase to which free LIGHTderived from the biological sample will be attached.

The solid phase to be used in the kits of the present inventionincludes, but is not limited, to microplates, magnetic particles, filterpapers for immunochromatography, polymers such as polystyrene, glassbeads, glass filters and other insoluble carriers. In one embodiment, asolid substrate containing many compartments or regions has at least onecompartment coated with antibodies of the invention.

The kits of the invention may also include a further component to thediagnostic agent, the anti-LIGHT antibody. The further component mayinclude, but is not limited, to enzymes for labeling, substratestherefor, radioisotopes, light-reflecting substances, fluorescentsubstances, colored substances, buffer solutions, and plates, and thosementioned hereinabove.

Free LIGHT Detection Assays

Disclosed are assays, which are methods for detecting and measuring freeLIGHT in a sample suspected to contain free LIGHT from a subject. Suchmethods may use antibody pairs for sandwich immunoassay in which freeLIGHT from a sample is captured by capture antibodies coating, e.g.,paramagnetic beads and a detection antibody further binds to thecaptured free LIGHT. A labeling enzyme conjugate such asstreptavidin-ß-galactosidase (SßG), is then added and binds to thedetection antibodies. Finally, an enzyme substrate is added to yieldfluorescence intensities that can correlate with the presence and amountof free LIGHT in the sample.

In some embodiments, a sample suspected of containing free LIGHT iscontacted with a capturing molecule for free LIGHT that specificallybinds to free LIGHT, but not to bound LIGHT, and is incubated to allowthe capturing molecule to bind to free LIGHT. Such embodiments furtherentail detecting the binding of free LIGHT to the capturing molecule anddetermining the amount of free LIGHT in the sample. The capturingmolecule may be, but need not be, an antibody. In some embodiments,bound LIGHT refers to LIGHT bound to a complex. In some embodiments,bound LIGHT is LIGHT bound to a complex with DcR3. In some embodiments,bound LIGHT is LIGHT that is neutralized or inactivated by DcR3. In someembodiments, bound LIGHT is LIGHT bound to either of its TNF receptors,HVEM and LTβR. In some embodiments, LIGHT bound to a complex comprisesLIGHT bound to DcR3 or an anti-LIGHT antibody.

In some embodiments, the capturing molecule specifically binds to freeLIGHT, but not to a LIGHT/DcR3 complex. In some embodiments, thecapturing molecule specifically binds to free LIGHT at the site at whichfree LIGHT binds to DcR3 or in the vicinity of the site at which freeLIGHT binds to DcR3 such that, for example, LIGHT cannot simultaneouslybind to the capturing molecule and to DcR3. In some embodiments, thecapturing molecule specifically binds to free LIGHT at a site at whichLIGHT binds to DcR3. In some embodiments, the capturing moleculespecifically binds to free LIGHT at the site or in the vicinity of thesite at which free LIGHT binds to either of its TNF receptors, HVEM andLTβR. In some embodiments, the capturing molecule specifically binds tofree LIGHT at the site at which free LIGHT binds to either of its TNFreceptors, HVEM and LTβR.

In some embodiments of the methods for detecting and measuring freeLIGHT in a sample suspected to contain free LIGHT from a subject, thesample is not only contacted with a capturing molecule, but also with adetection molecule. Such embodiments further entail incubating thesample to allow the capturing molecule and the detection molecule tobind to free LIGHT. Such embodiments further entail detecting thebinding of free LIGHT to the capturing molecule and determining theamount of free LIGHT in the sample. In some embodiments, the binding offree LIGHT to the detection molecule is also detected. In someembodiments, the detection molecule binds to a different site of freeLIGHT than the capture molecule.

In some embodiments of the methods for detecting and measuring freeLIGHT in a sample suspected to contain free LIGHT from a subject, themethods further entail comparing the amount of free and amount of totalLIGHT in the sample.

In some embodiments, the capturing molecule is an antibody. In someembodiments, the capturing molecule is an antibody chosen frommonoclonal, polyclonal, chimeric, single chain, bispecific orbi-effective, simianized, human, and humanized antibodies. In someembodiments, the capturing antibody is a monoclonal antibody. In someembodiments, the capturing antibody is bound to a support (e.g.,nanoparticle in Simoa platform). In some embodiments, the capturingantibody is ProSci RF16062.

In some embodiments, the detection molecule is an antibody. In someembodiments, the detection molecule is a monoclonal antibody. In someembodiments, the capturing antibody is ProSci RF16062. In someembodiments, the capturing antibody is Enzo ALX-804-841-C100. In someembodiments, the detection antibody is DCABH-13797. In some embodiments,the detection antibody is ProSci RF16062. In some embodiments, thecapturing antibody is ProSci RF16062 and the detection antibody isDCABH-13797. In some embodiments, the capturing antibody is EnzoALX-804-841-C100 and the detection antibody is ProSci RF16062.

In some embodiments, the sample is serum, plasma, saliva, or stool. Insome embodiments, the sample is a serum sample.

Example 1

A study was conducted in which serum from subjects with acute COVID-19infections was tested to determine the levels of free LIGHT and othercytokines/inflammatory markers during the acute phase of the illness.Whether there are differences in LIGHT levels and other inflammatorymarkers in healthy controls versus subjects with mild to severe diseasewith ALI/ARDS was determined. It was expected that subjects with severeCOVID-19 infection will have higher levels of serum free LIGHT and otherinflammatory markers compared to healthy controls or subjects with amild or moderate coronavirus infection without ARDS, and that,furthermore, these markers could be useful in identifying subjects whomight benefit from treatment with anti-LIGHT therapy. Given LIGHT's rolein mediating immune response, particularly its roles in inflammatorycell infiltration and T cell activation, these subjects were expected tobenefit from anti-LIGHT antibody therapy. Thus, the assay results wereexpected to provide the clinician with guidance as to which therapeuticagents are appropriate.

The study included 30 healthy controls, 28 subjects who were intubatedand had severe COVID-19 infection, and 20 subjects who werenon-ventilated and had mild to moderate COVID-19 infection. Blood wasdrawn from the subjects at a single timepoint. Using the free LIGHTassay described herein in Example 4, the presence and level of free(active) LIGHT in the blood serum was determined; the determination tookplace upon intubation for intubated patients.

Free LIGHT was significantly (p<0.0001) elevated in COVID-19 subjects(N=47) as compared to healthy controls (N=30). See FIG. 6 .

Free LIGHT was also significantly (p<0.0001) elevated in serum ofnon-ventilated COVID-19 patients (N=20) and intubated COVID-19 patients(N=27) as compared to healthy controls (N=30). See FIG. 7 .

Additional evidence suggests that the highest free LIGHT levels werefound in patients who required ventilator support, particularly inpatients over the age of 60, and that elevated free LIGHT levels areassociated with fatal outcomes in these patients, with a statisticallysignificant association between higher serum LIGHT levels and mortalityin patients over the age of 60 (p=0.0209). See FIG. 8 . The observedmortality rate was higher for patients over 60 years of age (82%)compared to patients <60 years (32%).

Example 2

A randomized, double-blind, placebo-controlled, multicenter, phase 2clinical trial to evaluate the efficacy and safety of an anti-LIGHTantibody in adults with COVID-19 pneumonia and acute lung injury wasconducted according to the methods described in Example 2. Theanti-LIGHT antibody has a VH of SEQ ID NO. 8, and a VL of SEQ ID NO: 9.

Example 2.1—Study Objectives and Endpoints

The primary objective of the study is to evaluate the effect of theanti-LIGHT monoclonal antibody compared with placebo in addition tostandard of care on prevention of ARDS in adults with COVID-19 pneumoniaand acute lung injury. The human anti-LIGHT monoclonal antibodyadministered comprises a heavy chain and a light chain that togethercomprise CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 amino acidsequences: SEQ ID NOs: 2, 3, 4, 5, 6, and 7.

The secondary objectives of the study are: to evaluate the safety andtolerability of the anti-LIGHT monoclonal antibody compared with placeboin addition to standard of care, in adults with COVID-19 pneumonia andacute lung injury; and to evaluate the effect of the anti-LIGHTmonoclonal antibody compared with placebo in addition to standard ofcare, on mortality in adults with COVID-19 pneumonia and acute lunginjury.

The exploratory objectives are: to evaluate the effect of the anti-LIGHTmonoclonal antibody compared with placebo in addition to standard ofcare, on viral load in adults with COVID-19 pneumonia and acute lunginjury; and to evaluate the PK, pharmacodynamics (PD), andimmunogenicity of the anti-LIGHT monoclonal antibody in adults withCOVID-19 pneumonia and acute lung injury.

The primary endpoint of the study is: the proportion of subjects aliveand free of respiratory failure over 28 days. Respiratory failure isdefined based on resource utilization including one of the following:endotracheal intubation and mechanical ventilation; oxygen delivered byhigh-flow nasal cannula (heated, humidified, oxygen delivered viareinforced nasal cannula at flow rates >20 L/min with fraction ofdelivered oxygen ≥0.5); noninvasive positive pressure ventilation; orextracorporeal membrane oxygenation (ECMO).

The secondary endpoints of the study are: 1-month mortality defined asthe proportion of subjects who were alive at the Day 28/earlytermination (ET); partial pressure of arterial oxygen/percentage ofinspired oxygen (PaO2/FiO2) ratio; time to invasive ventilation;duration of ventilation support; intensive care unit (ICU) length ofstudy; hospital length of stay; time to return to room air with restingpulse oximeter >93%; peak PaO2/FiO2 ratio; partial pressure of oxygen(P02); change in Sequential Organ Failure Assessment (SOFA) score;change in body temperature; and adverse event (AE) monitoring and safetylaboratory determination.

The exploratory endpoints of the study are viral load in nasopharyngealaspirates, LIGHT levels and inflammatory biomarker patterns(InflammationMAP), plasma concentrations of the anti-LIGHT monoclonalantibody over time, and measurement of anti-drug antibody (ADA).

Example 2.2—Study Design

This study is designed to determine the efficacy and safety of theanti-LIGHT monoclonal antibody compared with placebo in addition tostandard of care, administered SC in subjects with COVID-19 pneumoniaand acute lung injury. The primary objective of the study is to evaluatethe effect of the anti-LIGHT monoclonal antibody on prevention of ARDSin adults with COVID-19 pneumonia and acute lung injury. The efficacy ofthe anti-LIGHT monoclonal antibody is determined by measuring PaO2/FiO2;alive, respiratory failure days; alive ventilator-free days; ICU andhospital length of stay; return to room air or baseline oxygenrequirement; SOFA score; body temperature; viral load in nasopharyngealaspirates; time to invasive ventilation; and duration of ventilationsupport. These are routinely used and accepted methods to assessrespiratory functions. The study also assesses 1-month mortality rate,apart from other parameters.

The study takes place at approximately 10 study sites in the US.

83 patients are enrolled in the study. A screening log of studycandidates is maintained at each study site.

Inclusion Criteria

Subjects fulfill the following requirements to be randomized into thestudy:

1. Subject/legally authorized representative (LAR) is able to understandand provide written informed consent and assent (as applicable) toparticipate in this study.

2. Subject is ≥18 years of age at the time of informed consent andassent (as applicable).

3. Subject is male or non-pregnant, non-lactating female, who if ofchildbearing potential agrees to comply with any applicablecontraceptive requirements if discharged from the hospital prior tocompleting the study.

4. Subject has a diagnosis of COVID-19 infection through an approvedtesting method.

5. Subject is hospitalized due to clinical diagnosis of pneumonia withacute lung injury defined as diffuse bilateral radiographic infiltrateswith PaO2/FiO2>100 and <300.

6. If available, subject's oxygen saturation at rest in ambient air<93%.

Exclusion Criteria

The presence of any of the following criteria excludes a subject fromthe study:

1. Subject is intubated.

2. Subject is taking immunomodulators or anti-rejection drugs. The useof corticosteroids as part of standard of care measures in severeCOVID-19 patients is permitted when clinically indicated and discussedwith the Medical Monitor.

3. Subject has been administered an immunomodulating biologic drugwithin 60 days of baseline.

4. Subject is in septic shock defined as persistent hypotensionrequiring vasopressors to maintain mean arterial pressure (MAP) of 65 mmHg or higher and a serum lactate level greater than 2 mmol/L (18 mg/dL)despite adequate volume resuscitation.

5. Subject has ALT/AST >5×ULN or creatinine >2.5 mg/dl

6. Subject has neutrophils <500/ml3

7. Subject has platelets <50,000/ml3

8. Subject has known hypersensitivity to any of the components of theanti-LIGHT monoclonal antibody

9. Subject has received any live attenuated vaccine, such asvaricella-zoster, oral polio, or rubella, within 3 months prior to thebaseline visit.

Screen Failures

Subjects who fail inclusion and/or exclusion criteria are not rescreenedfor the study.

Premature Subject Withdrawal

All subjects are advised that they are free to withdraw fromparticipation in this study at any time, for any reason, and withoutprejudice. Investigators are instructed to make every reasonable attemptto keep subjects in the study; however, subjects have to be withdrawnfrom the study if they withdraw consent to participate.

The sponsor reserves the right to request the withdrawal of a subjectdue to protocol deviations or other reasons.

The investigator also has the right to withdraw subjects from the studyat any time for any reason. If a subject is withdrawn before completingthe study, the subject is supposed to be followed-up as instructed inthe Schedule of Assessments (Table 1). The reason for withdrawal has tobe determined by the investigator and recorded in the subject's medicalrecord and on the case report form (CRF). If a subject is withdrawn formore than 1 reason, each reason is supposed to be documented in thesource document and the most clinically relevant reason is to be enteredon the CRF.

Possible reasons for discontinuation include but are not limited to:

-   -   Adverse event    -   Major protocol deviation    -   Withdrawal by subject from study assessments    -   Withdrawal by subject from study drug    -   Lost to follow-up    -   Other. If Other selected, the investigator required to specify        the reason on the CRF.        Subject Replacement Criteria

Subjects who withdraw or are discontinued from the study are allowed tobe replaced.

TABLE 1 Schedule of Assessments Safety Follow-up Phone Baseline Day DayDay Day Day Day CallDay Procedure (Day 1) 2 5 8 9 14 28/ET¹ 60² Informedconsent and X assent (as applicable) Randomization X Concomitantmedications³ X X X X X X X X Adverse events⁴ X X X X X X X X Viral loadin X X nasopharyngeal aspirates⁹ ECG⁷ X X X X X X X Pregnancy test¹⁰ XPK  X⁶ X X X LIGHT and  X⁵  X⁶ X X X X X InflammationMAP ADA⁸ X X XInvestigational product X administration Abbreviations: ADA = anti drugantibodies; EC = electrocardiogram; ET = early termination;InflammationMAP = inflammatory biomarker patterns; LIGHT =Lymphotoxin-like, exhibits Inducible expression, and competes withHerpes Virus Glycoprotein D for Herpesvirus Entry Mediator, a receptorexpressed by T lymphocytes; PK = pharmacokinetics. ¹Day 28 is conductedas a follow-up call for subjects who are discontinued from the study andfor subjects who are discharged prior to the Day 28 visit. Additionally,if a subject is discontinued from the study or discharged from thehospital prior to Day 28, the Day 28/ET visit procedures are to beperformed. No further visits are required to be performed with theexception of the Day 28 and Day 60 follow-up calls. ²A safety follow-upcall is conducted approximately 59 days (±7 days) after administrationof investigational product. ³Concomitant medications are collectedthroughout the study period. ⁴Adverse events are collected throughoutthe study period. ⁵Samples ae collected prior to investigational productadministration. ⁶Samples are collected 24 hours (±2 hours) post Day 1dosing. ⁷An ECG is collected daily when a subject is not being assessedby cardiac monitoring. ⁸An ADA sample is collected at Days 8, 14, 28/ET.Additionally, a sample is collected when an immunologically relatedadverse event is reported (e.g., a skin reaction, lupus-like syndrome,unexplained thrombocytopenia). ⁹If the site is unable to do aquantitative test in order to obtain viral load, the viral load samplesto be collected at Days 1 and 5 are not required to be collected. ¹⁰Forfemales of childbearing potential. A subject is not considered to be ofchildbearing potential if they are post-menopausal (12 consecutivemonths of spontaneous amenorrhea and ≥ age 51 years, and/or surgicallysterile (having undergone one of the following surgical acts:hysterectomy, bilateral tubal ligation, bilateral oophorectomy orbilateral salpingectomy) and at least 6 weeks post-sterilization.

TABLE 2 Standard of Care Procedures Procedure ABG Chest CT/CXR GlasgowComa Scale Laboratory procedures including CBC, chemistry and urinalysisCRP Physical examination Pulse Oximetry SOFA Temperature Vital signsAbbreviations: ABG = arterial blood gas; CBC = complete blood count; CRP= C-reactive protein; CT = computed tomography; CXR = chest x-ray; SOFA= Sequential Organ Failure Assessment. These procedures are performedper the site's practice unless defined otherwise

Example 2.3—Treatments

Identification of Investigational Product(s), Dose and Mode ofAdministration

The anti-LIGHT monoclonal antibody is the investigational product thatis used in this study. It is supplied in vials. Placebo is sourcedlocally and provided as volume-matched normal saline for injection. Theanti-LIGHT monoclonal antibody or placebo is administered by SCinjection in the abdomen in a zone of 4 to 10 cm from the umbilicus withthe injection site rotated based on the number of syringes used. Theanti-LIGHT monoclonal antibody or placebo is administered at baseline(Day 1). The anti-LIGHT monoclonal antibody is administered at 16 mg/kgdose (maximum dose of 1200 mg).

Labeling and Packaging

All packaging and labeling operations are performed by the sponsor ordesignee per Good Manufacturing Practice and Good Clinical Practice(GCP) rules. The investigational product is sent to the study site bythe sponsor or designee. Labeling is in local language and dependentupon local regulations.

Treatments Administered

Eligible subjects receive the anti-LIGHT monoclonal antibody or placeboon Day 1 in addition to standard of care. The standard of care is to bemaintained throughout the study and is allowed to include off-label useof other drugs, devices, or interventions used to treat COVID-19.

Dispensing and Storage

The anti-LIGHT monoclonal antibody supplied by sponsor is usedexclusively in this clinical study per the instructions of the protocol.Placebo is sourced locally and provided as volume matched injection ofnormal saline. The investigator is responsible for dispensing theinvestigational product per the dosage scheme and for ensuring properstorage of the investigational product.

The unblinded pharmacist is to confirm the receipt of theinvestigational product with his/her signature. A copy of this receiptis to be kept by the unblinded pharmacist, and another copy is stored atsponsor and/or designee. Until the investigational product is dispensedto the subjects, it is stored at 2° C. to 8° C. (35.6° F. to 46.4° F.)and protected from light. Investigators or other authorized persons(e.g., pharmacists) are responsible for storing the investigationalproduct provided by the sponsor in a secure and safe place in accordancewith local regulations, labeling specifications, institutional policiesand procedures.

Control of storage conditions for the investigational product providedby the sponsor, especially control of temperature (e.g., refrigeratedstorage) and daily temperature monitoring, and information on in-usestability and instructions for handling the investigational product areto be managed according to the rules provided by the sponsor.

Blinding and Unblinding Treatment Assignment

This is a double-blind study. All subjects, investigators, and studypersonnel involved in the conduct of the study, including datamanagement, are blinded to treatment assignment except for the followingindividuals: specified unblinded statistician from the Contract ResearchOrganization (CRO) who has access to the randomization code; andspecified unblinded pharmacist(s) from the hospital who have access tothe randomization code in order to prepare the investigational product.

The unblinded pharmacist(s) and unblinded statistician do not otherwiseparticipate in the study or data analysis prior to unblinding of thestudy.

Treatment unblinding is discouraged if knowledge of the treatmentassignment does not materially change the planned management of amedical emergency. Unblinding is permitted in a medical emergency thatrequires immediate knowledge of the subject's treatment assignment.Whenever possible unblinding is to be discussed with the Sponsor MedicalMonitor. For emergency unblinding the Investigator is to contact theunblinded pharmacist(s). If the Investigator is not able to discusstreatment unblinding in advance, then they are to notify the SponsorMedical Monitor as soon as possible about the unblinding incidentwithout revealing the subject's treatment assignment. The Investigatoror designee is to record the date and reason for study discontinuationon the appropriate eCRF for that subject. In all cases that are notemergencies, the Investigator is to discuss the event with the SponsorMedical Monitor prior to unblinding the subject's treatment assignment.

If the treatment assignment is unblinded for an individual subject, theInvestigator is to be notified of that subject's treatment assignmentwithout unblinding the treatment assignments for the remaining subjectsin the study. The Investigator is to make this decision afterconsultation with the Sponsor Medical Monitor.

Selection of Doses in the Study

The dose of 16 mg/kg (maximum dose of 1200 mg) is selected in an effortto maximize the ability to achieve the blockade of LIGHT while ensuringpatient safety. In a robust toxicology program the anti-LIGHT monoclonalantibody was dosed as high as 100 mg/kg in monkeys and was welltolerated while the NOAEL was determined to be 60 mg/kg. In dosing inhumans, the anti-LIGHT monoclonal antibody was safe and well toleratedin single ascending doses up to 1200 mg in healthy volunteers. Therewere no clinically meaningful treatment-emergent adverse events orchanges in ECG parameters or laboratory values. A safety reviewcommittee analyzed data from individual subjects and across all subjectstreated in a daily fashion to assess any safety signals with dosing.

Dose Adjustment Criteria

No dose adjustments are allowed.

Drug Accountability

Records showing the receipt, dispensing, or other disposition of theinvestigational product including the date, lot identifier, dosage,volume administered to each subject, and identification of subjects(subject number and initials) who receive the investigational productare maintained by an unblinded pharmacist. The investigator does notsupply the investigational product to any person except those named assubinvestigators on the US Food and Drug Administration (FDA) Form FDA1572 and designated study personnel, and subjects in this study. Theinvestigator does not dispense the investigational product from anystudy locations other than those listed on Form FDA 1572. If any of theinvestigational product is not dispensed, is lost, stolen, spilled,unusable, or received in a damaged container, that information is to bedocumented and reported to sponsor and appropriate regulatory agencies,as required.

Upon completion of the study, unused investigational product may be leftin the original packaging for final disposition by the sponsor or perthe site's standard practice. Any partially used investigational productand all empty packaging (e.g., vials) may be saved for final dispositionby the sponsor, returned to the sponsor's designee for destruction, orper the site's standard practice.

Permitted and Prohibited Therapies

All non-study therapies including but not limited to over-the-counterand non-pharmacological treatments received within 7 days prior tobaseline and through the end of study are recorded on the appropriateelectronic case report form (eCRF) page.

Prior Therapies

Prior treatment includes all treatment received within 7 days of thedate of first dose of investigational product. Prior treatmentinformation is recorded on the appropriate eCRF page.

Concomitant Therapies

Concomitant therapies refer to all therapies taken between the dates ofthe first dose of investigational product and the end of the follow-upperiod, inclusive. Concomitant treatment information is recorded on theappropriate CRF page.

Permitted Therapies

Medications considered necessary for the subject's welfare areadministered at the discretion of the investigator. The Sponsor MedicalMonitor may be contacted in the event the site is in a situation wherefurther clarity is needed.

Acceptable methods of birth control are implants, injectables, combinedoral contraceptives, intrauterine device, sexual abstinence orvasectomized partner.

Prohibited Therapies

Subjects may not have been administered an immunomodulating biologicdrug within 60 days prior to the baseline visit, or have received anylive attenuated vaccine, such as varicella-zoster, oral polio, orrubella, within 3 months prior to the baseline visit. During the study,new initiation of investigational compounds may be prohibited, with theexception of corticosteroids, which are permitted to be administered aspart of standard of care measures in severe COVID-19 patients whenclinically indicated and discussed with the Medical Monitor. The use ofimmunomodulatory or anti-rejection drugs may be prohibited.

Treatment after End of Study

Subjects are treated per standard clinical practice throughout thestudy.

Example 2.4—Study Procedures

Subjects/LAR provide written informed consent and assent (as applicable)before study-related procedures are initiated.

For the timing of assessments and procedures throughout the study, referto the schedule of events (Table 1 and Table 2). The procedures listedin Table 2 are considered standard of care and are performed per thesite's practice unless defined otherwise. Throughout the study, everyreasonable effort is made by study personnel to follow the timing ofassessments and procedures in the schedule of events for each subject.

Study Duration

The anti-LIGHT monoclonal antibody and placebo treatment areadministered on Day 1 and the duration of the study period was 60 days.

Efficacy Assessments

Efficacy response is assessed by the following procedures at the timepoints mentioned in the Schedule of Assessments (Table 1 and Table 2):Arterial blood gas (ABG) test to measure PaO2/FiO2; pulse oximetry; theSOFA score measurement (The SOFA score is a simple and objective tool tocalculate both the number and the severity of organ dysfunction in thefollowing 6 organ systems: respiratory, coagulatory, liver,cardiovascular, renal, and neurologic [Table 3], and the score canmeasure individual or aggregate organ dysfunction (Vincent J L, MorenoR, Takala J, et al. The SOFA (Sepsis-related Organ Failure Assessment)score to describe organ dysfunction/failure. On behalf of the WorkingGroup on Sepsis-Related Problems of the European Society of IntensiveCare Medicine. Intensive Care Med. 1996; 22(7):707-710]); bodytemperature; viral load in nasopharyngeal aspirates. Apart from thesetests, time to invasive ventilation and duration of ventilation support,are also recorded.

TABLE 3 The Sequential Organ Failure Assessment (SOFA) score SOFA Score1 2 3 4 Respiration PaO2/FiO2 <400 <300 <200 (with <100 (with (mmHg)respiratory respiratory support) support) Coagulation Plate- <150 <100<50 <20 lets ×10³/mm³ Liver Bilirubin 1.2-1.9 2.0-5.9  6.0-11.9   >12.0(mg/dL) Cardiovascular^(a) Hypotension MAP <70 Dopa- Dopamine >5Dopamine >15 mmHg mine ≤5 or epineph- or epineph- ordobuta- rine ≤0.1rine >0.1 mine or norepi- or norepi- (any dose) nephrine ≤0.1nephrine >0.1 Central Nervous System Glasgow 13-14 10-12 6-9  <6 ComaScore Renal Creatinine 1.2-1.9 2.0-3.4 3.5-4.9 or <500 >5.0 or <200(mg/dL) orurine output (mL/day) MAP: mean arterial pressure; PaO2/FiO2:partial pressure of arterial oxygen/percentage of inspired oxygen.^(a)Adrenergic agents administered for at least 1 h (doses given are inμg/kg-min). Reference: Vincent JL, Moreno R, Takala J, et al. The SOFA(Sepsis-related Organ Failure Assessment) score to describe organdysfunction/failure. On behalf of the Working Group on Sepsis-RelatedProblems of the European Society of Intensive Care Medicine. IntensiveCare Med. 1996; 22(7): 707-710.Safety Assessments

Safety and tolerability assessments include the frequency and severityof AEs as well as the evaluation of changes in clinical laboratoryvalues, vital signs, ECG recordings, and physical examination findings.

Clinical Laboratory Tests to be Performed

A sample for C-reactive protein is collected at the time pointsspecified in the Schedule of Assessments (Table 1). With the exceptionof PK, LIGHT and InflammationMAP samples, all other laboratory samplesare considered standard of care and may be performed per the site'spractice.

Laboratory specimens are analyzed at the hospital laboratory per theircollection and processing requirements.

Sampled Blood Volume

The sampled blood volume for this study is shown in Table 4.

TABLE 4 Sampled Blood Volume per Subject Sample Total Volume Number ofVolume Assessment (mL) Samples (mL) Anti-LIGHT 2.0 4 8.0 monoclonalantibody concentration and PK analysis Anti-drug Antibodies 2.0 3 6.0LIGHT/InflammationMAP 2.5 7 17.5 Total mL — — 27.5 InflammationMAP =inflammatory biomarker patterns; LIGHT = Lymphotoxin-like, exhibitsInducible expression, and competes with Herpes Virus Glycoprotein D forHerpesvirus Entry Mediator, a receptor expressed by T lymphocytesAnti-LIGHT Monoclonal Antibody Concentration, Pharmacokinetic andAnti-Drug Antibody Assessments

Pharmacokinetics and ADA assessments are calculated from the plasmaconcentrations of the anti-LIGHT monoclonal antibody.

Specimen Handling Requirements

The transmission of infectious agents may occur through contact withcontaminated needles and blood or blood products. Consequently,appropriate blood and body fluid precautions may be employed by allstudy personnel involved in the collection of blood and handling ofspecimens in both the clinic and laboratory settings. Refer to currentrecommendations of the appropriate authorities.

In addition to appropriate handling of subject samples, specificregulations exist regarding the shipment of biologic/etiologic samples.Procedures and regulations for the packaging and shipping of infectioussamples are outlined in the site and/or study laboratory manual. Theinvestigator is responsible for ensuring that all study samples that aretransported to another location are appropriately packed and shipped perthe applicable regulations.

Evaluation of Laboratory Values

The normal ranges of values for the laboratory assessments in this studyare provided by the local laboratory of each hospital. They are regardedas the reference ranges on which decisions are be made for the specificsite.

If a laboratory value is out of the reference range, it is notnecessarily clinically relevant. The investigator may evaluate theout-of-range values and record his/her assessment of the clinicalrelevance in the subject's source documentation.

All laboratory values which, in the investigator's opinion, showclinically relevant or pathological changes during or after terminationof the treatment are to be discussed with the medical monitor, asnecessary, and reported as AEs and followed.

All measurements described in this section are recognized standardmethods.

Clinical Examination: Blood Pressure, Pulse Rate, Respiratory Rate,Temperature, Height, and Body Weight

Blood pressure, pulse rate, respiratory rate, temperature, height, andbody weight are considered standard of care and are performed per thesite's practice. Additional blood pressure and pulse rate measurementsare allowed to be performed, as determined by the investigator, toensure appropriate monitoring of subject safety and accurate recordingof vital sign measurements. Any changes from baseline which are deemedclinically significant by the investigator are recorded as an AE.

Clinical Examination: Electrocardiogram

A standard 12-lead ECG is considered standard of care and is performeddaily for those who are not being assessed by cardiac monitor. They areperformed per the site's practice unless defined otherwise in Table 2.All ECG recordings are identified with the subject number, subjectinitials, date, and time of the recording and a copy is included withthe subject's source documentation. All ECGs may be performed using theequipment supplied by the investigational site.

Electronic ECG tracings are analyzed per the site's practice. Inaddition, the investigator's assessment of the ECG tracing as normal orabnormal are documented, and if abnormal, his/her determination ofwhether the abnormality is clinically significant or not may bedocumented on the tracing.

All ECG values which, in the investigator's opinion, show clinicallyrelevant or pathological changes during or after termination of thetreatment may be discussed with the Sponsor Medical Monitor and reportedas AEs and followed.

Clinical Examination: Physical Examination

A complete physical examination is considered standard of care and isperformed per the site's practice unless defined otherwise in Table 2.Any clinically significant physical examination findings are reported asAEs and followed.

Clinical Examination: Adverse Events

The investigator is responsible for the detection and documentation ofevents meeting the criteria and definition of an AE or SAE describedpreviously. Any clinically relevant observations made during the periodof hospitalization are considered AEs.

Pharmacokinetics and Immunogenicity Analyses

Blood samples for PK analysis are collected on Day 2 at 24 hours (±2hours) post dose and at any time on Days 8, 14 and 28. Blood samples forADA analysis are collected at any time on Days 8, 14 and 28.Additionally, a sample is collected when an immunologically relatedadverse event is reported. Samples are processed to plasma. Time of PKsamples is recorded in the eCRF. A total of 1.0 mL plasma per PK and ADAsample are collected from each subject to measure plasma concentrationsof the anti-LIGHT monoclonal antibody and ADAs. Pharmacokinetic and ADAsamples are to be processed according to the methods and directions setforward in the Laboratory Manual(s) and guidance(s). Pharmacokinetic andADA plasma sample analysis are performed by laboratory defined in theLaboratory Manual(s) and guidance(s), according to their standardoperating procedures (SOPs) using a validated enzyme-linkedimmunosorbent assay (ELISA). Assay and analysis details are described inthe method validation and bioanalytical information.

Pharmacodynamics

Blood samples are collected for exploratory analyses. Exploratoryanalyses include are not limited to LIGHT levels and InflammationMAP asspecified in Table 1. Exploratory biomarker analyses are performed atthe laboratories specified in the Laboratory Manual(s) and guidance(s).

Example 2.5—Adverse Events

Adverse Event Collection

An AE is defined as any untoward medical occurrence in a patient orclinical investigation subject administered a pharmaceutical productthat does not necessarily have a causal relationship with the product.An AE could therefore be any unfavorable and unintended sign (includinga new, clinically important abnormal laboratory finding), symptom, ordisease, temporally associated with the product, whether related to theproduct. An AE is considered treatment-emergent if it occurred after thefirst dose of investigational product and within 30 days of a subject'slast dose of investigational product.

All AEs are collected from the time the informed consent was signeduntil the end of study (Day 60). This includes events occurringregardless of whether investigational product is administered. Wherepossible, a diagnosis rather than a list of symptoms is recorded. If adiagnosis is not yet made, then each symptom is listed individually. AllAEs are captured on the appropriate AE pages in the eCRF and in sourcedocuments. In addition, to untoward AEs, unexpected benefits outside theinvestigational product indication are captured in the source documentsand AE eCRF.

All AEs are followed to closure (the subject's health has returned tohis/her baseline status or all variable have returned to normal),regardless of whether the subject was still participating in the study.Closure indicates that an outcome has been reached, stabilizationachieved (the investigator does not expect any further improvement orworsening of the event), or the event was otherwise explained. Whenappropriate, medical tests and examinations are performed so thatresolution of an event(s) could be documented.

Severity of Adverse Events

The severity of AEs are recorded during the course of the eventincluding the start and stop dates for each change in severity. An eventthat changes in severity is captured as a new event. Worsening of apre-treatment events, after initiation of investigational product isrecorded as new AEs. For example, if the subject experiences mild,intermittent headaches prior to dosing with investigational product;however, the headache intensity increases to moderate after the firstdose of investigational product, a new AE of moderate intermittentheadaches is recorded in the source documents and eCRF.

The medical assessment of clinical severity of an AE is determined usingthe definitions outlined in Common Terminology Criteria for AdverseEvents (CTCAE), Version 5.0 (Published Nov. 27, 2017 by the USDepartment of Health and Human Services, National Institutes of Health,National Cancer Institute). Grade 1 is defined as mild; asymptomatic ormild symptoms; or clinical or diagnostic observations only; orintervention not indicated. Grade 2 is defined as moderate; or minimal,local or non-invasive intervention indicated; or limitingage-appropriate instrumental activities of daily living (ADL). Grade 3is defined as severe or medically significant but not immediatelylife-threatening; or hospitalization or prolongation of hospitalizationindicated; or disabling; or limiting self-care ADL. Grade 4 is definedas life-threatening consequences; or urgent intervention indicated.Grade 5 is defined as deaths related to AE.

The above-referenced CTCAE document may be referred to for fulldescription of CTCAE terms and instrumental and self-care ADLs. Severityis a classification of intensity whereas an SAE is an AE that meetsserious criteria.

Relationship Categorization

A physician investigator makes the assessment of relationship toinvestigational product for each AE. The investigator decides whether,in his or her medical judgment, there is a reasonable possibility thatthe event may have been caused by the investigational product. If thereis no valid reason for suggesting a relationship, then the AE may beclassified as “not related”.

Otherwise, the AE is categorized per the guidelines below. The causalityassessment may be documented in the source document and the eCRF (Table5).

TABLE 5 Assessment of Relationship to Investigational ProductRelationship Description Not Related Exposure to investigational producthas not occurred. OR The administration of investigational product andthe occurrence of the AE are not reasonably related in time OR The AE isconsidered likely to be related to an etiology other than the use of theinvestigational product, that is, there are no facts/evidence orarguments to suggest a causal relationship to the investigationalproduct. Possibly The administration of the investigational product andthe Related occurrence of the AE are reasonably related in time. AND TheAE could not be explained equally well by factors or causes other thanexposure to investigational product Probably The administration ofinvestigational product and the Related occurrence of the AE arereasonably related in time. AND The AE is more likely explained byexposure to investigational product than by other factors or causes.Outcome at the Time of Last Observation

The outcome at the time of last observation is classified as:recovered/resolved; recovered/resolved with sequelae;recovering/resolving; not recovered/not resolved; fatal; or unknown.

Reporting of Serious Adverse Events

Initial and follow-up SAE reports are completed by the investigator ordesignee and sent to the CRO within 24 hours of the first awareness ofan SAE. The investigator or designee completes, signs and dates theappropriate SAE form and verifies the accuracy of the informationagainst corresponding source documents. This information is sent to theCRO Pharmacovigilance Department.

Serious Adverse Event Definition

An SAE is defined as any untoward medical occurrence, whether consideredto be related to investigational product or not, that at any dose:results in death; is life-threatening; requires inpatienthospitalization or prolongation of existing hospitalization; results inpersistent or significant disability/incapacity; is a congenitalanomaly; or is an important medical event.

Note that the term “life-threatening” in the definition of “serious”refers to an event in which the subject is at risk of death at the timeof the event; it does not refer to an event which hypothetically mighthave caused death if it were more severe.

Note that the term “inpatient hospitalization” is defined as 24 hours ina hospital or an overnight stay. An elective hospital admission to treata condition present before exposure to the test drug, or a hospitaladmission for a diagnostic evaluation of an AE, does not qualify thecondition or event as an SAE. Further, an overnight stay in the hospitalthat is only due to transportation, organization, or accommodationproblems and without medical background does not need to be consideredan SAE.

Note that the term “congenital anomaly” refers to an infant born to amother who was exposed to the investigational product during pregnancyis an SAE. However, a newly diagnosed pregnancy in a subject thatreceives an investigational product is not to be considered an SAEunless it is suspected that the investigational product(s) interactedwith a contraceptive method and led to the pregnancy.

Note that medical and scientific judgment may be exercised in decidingwhether it is appropriate to consider other situations serious, such asimportant medical events that are not immediately life-threatening or donot result in death or hospitalization but may jeopardize the subject ormay have required intervention to prevent one of the other outcomeslisted in the definition above. Examples of such events are intensivetreatment in an emergency room or at home for allergic bronchospasm,blood dyscrasias or convulsions that do not result in hospitalization,or development of drug dependency or drug abuse.

Serious Adverse Event Collection Time Frame

All SAEs, regardless of the relationship to study, are collected fromthe time the subject/LAR signed the informed consent and assent [ifapplicable] until the subject's last contact. The investigator ordesignee was to report all SAEs promptly to CRO within 24 hours of firstbecoming aware of the event.

Any SAE(s), regardless of relationship to study, discovered by theinvestigator at any interval after study was completed are reported toCRO within 24 hours of the first awareness of the event.

Serious Adverse Event Onset and Resolution Dates

The onset date of the SAE is defined as the date the event meets seriouscriteria. The resolution date is the date the event no longer meetsserious criteria, the date symptoms resolve, or the event is consideredchronic. In the case of hospitalization, the hospital admission anddischarge dates are considered respectively, the onset and resolutiondate of the SAE.

Any signs or symptoms experienced by the subject after signing theinformed consent form and assent form (if applicable), or leading up tothe onset date of the SAE or following the resolution date of the SAEare recorded as an AE.

Fatal Outcome

Fatal is only selected as an outcome when the AE results in death. Ifmore than 1 AE is possibly related to the subject's death, the outcomeof death is to be indicated for each such AE.

Any AE that results in the subject's death have fatal checked as anoutcome with the date of death recorded as the resolution date. AEsresulting in death were required to be reported within 24 hours as aSAE, if not already reported as such. In the event of a subject's death,data was to be collected on whether the death occurred after thewithdrawal of care and, if so, the reason for the withdrawal of care.

For other AEs, ongoing at the time of death that did not contribute tothe subject's death, the outcome was to be considered not resolved,without a resolution date recorded.

Adverse Events of Special Interest

There are no events from research to date which qualify as AEs ofspecial interest.

Adverse drug reactions observed in an anti-LIGHT monoclonal antibodypre-clinical observation include injection site reactions.

Observations for reactions with other biological agents include:potential for increased infection (including opportunistic infectionssuch as tuberculosis); hypersensitivity reactions (includinganaphylaxis); immunogenicity; malignancy; and impaired immunization.

Any new infection that occurred on study, regardless of the infectingagent (i.e. viral or non-viral), was to be captured. Additionally, thesite of infection and source of culture (bronchoalveolar lavage,tracheal aspirate, sputum, blood, urine etc.) was to be captured.

Pregnancy

All females of childbearing potential who participate in the study arecounseled on the need to practice adequate birth control and on theimportance of avoiding pregnancy during study participation. Females areinstructed to contact the investigator or study staff immediately ifpregnancy occurred or is suspected.

Pregnancy testing is conducted on females of childbearing potential atbaseline. Any female who is found to be pregnant at baseline is excludedfrom the study and considered to be a screening failure. Any female whois found to be pregnant after the dosing is required to be discontinuedfrom the study and the end of study visit assessments performed as soonas possible after learning of the pregnancy.

The investigator reports the pregnancy of any female (study participantor female partner of male study participant) who became pregnant duringinvestigational product treatment or within 60 days of being randomizedand receiving the investigational product. The pregnancy is reportedwithin 24 hours of learning of the pregnancy to the CRO using thePregnancy Data Collection Form via the same fax and email address as forSAE reporting. The investigator contacts the designated individual(s)who receive SAE notification and record information related to thepregnancy on an Exposure in Utero form/other designated form provided bythe sponsor or its designee.

The investigator is also responsible for following the pregnancy untildelivery or termination. These findings are reported on the PregnancyData Collection Form and forwarded to the designated individual(s). Theevent meets the SAE criterion only if it results in a spontaneousabortion or a congenital anomaly.

Anaphylaxis

Any AE that represents an anaphylactic reaction is classified using thedefinitions provided in Sampson H A, Munoz-Furlong A, Campbell R L, etal. Second symposium on the definition and management of anaphylaxis:Summary Report-Second National Institute of Allergy and InfectiousDisease/Food Allergy and Anaphylaxis Network Symposium. J Allergy ClinImmunol 2006; 117(2):391-97, and shown in Table 6.

TABLE 6 Clinical Criteria for Diagnosing Anaphylaxis Anaphylaxis ishighly likely when any one of the following 3 criteria are fulfilled: 1.Acute onset of an illness (minutes to several hours) with involvement ofthe skin, mucosal tissue, or both (eg, generalized hives, pruritus orflushing, swollen lips-tongue-uvula) AND AT LEAST ONE OF THE FOLLOWINGa. Respiratory compromise (eg, dyspnea, wheeze-bronchospasm, stridor,reduced PEF, hypoxemia) b. Reduced BP or associated symptoms ofend-organ dysfunction (eg, hypotonia [collapse], syncope, incontinence)2. Two or more of the following that occur rapidly after exposure to alikely allergen for that patient (minutes to several hours): a.Involvement of the skin-mucosal tissue (eg, generalized hives, itch-flush, swollen lips-tongue-uvula) b. Respiratory compromise (eg,dyspnea, wheeze-bronchospasm, stridor, reduced PEF, hypoxemia) c.Reduced BP or associated symptoms (eg, hypotonia [collapse], syncope,incontinence) 3. Reduced BP after exposure to known allergen for thatpatient (minutes to several hours): a. Infants and children: lowsystolic BP (age specific) or greater than 30% decrease in systolic BP*b. Adults: systolic BP of less than 90 mm Hg or greater than 30%decrease from that person's baseline *Low systolic blood pressure forchildren is defined as less than 70 mm Hg from 1 month to 1 year, lessthan(70 mm Hg + [2 × age]) from 1 to 10 years, and less than 90 mm Hgfrom 11 to 17 years. Abbreviations: Peak expiratory flow; BP = bloodpressure Reference: Sampson HA, Munoz-Furlong A, Campbell RL, et al.Second symposium on the definition and management of anaphylaxis:Summary Report-Second National Institute of Allergy and InfectiousDisease/Food Allergy and Anaphylaxis Network Symposium. J Allergy ClinImmunol 2006; 117(2): 391-97).

Example 2.6—Statistics

Sample Size Determination

A total of 83 subjects are randomized to one of two treatment groups(the anti-LIGHT monoclonal antibody or placebo in addition to standardof care) in a 1:1 ratio. One subject withdrew, leaving 82 subjects. Thissample size of 82 subjects provides greater than 80% power to detect adifference of 0.25 (25%) in the proportion of subjects alive and free ofrespiratory failure using a Chi-square exact test at a one-sidedsignificance level of 0.05. This calculation assumes that the proportionalive and free of respiratory failure is 0.60 in the placebo group and0.85 in the anti-LIGHT monoclonal antibody group.

Analysis Populations

This study has several populations of interest. The Randomized AnalysisSet includes all subjects who are randomized in the study. Subjects arecategorized according to their randomized treatment group. TheRandomized Analysis Set is used for all disposition, protocoldeviations, and demographic and other baseline characteristics analyses.The Safety Analysis Set includes all subjects who are randomized in thestudy and receive at least one dose of investigational product. Subjectsare categorized according to their actual treatment group. The SafetyAnalysis Set is used for all exposure and safety analyses. The FullAnalysis Set includes all subjects who receive at least one dose ofinvestigational product and have a baseline and at least onepost-baseline efficacy assessment. Subjects are categorized according totheir randomized treatment group. The Full Analysis Set is used for allefficacy and pharmacodynamic analyses. The PK Analysis Set includes allsubjects who receive at least one dose of investigational product andhave at least one post dose measurable plasma sample. Subjects arecategorized according to their actual treatment group. The PK AnalysisSet is used for all PK analyses.

Statistical Analyses

All efficacy and safety variables are summarized using descriptivestatistics. Descriptive statistics for continuous data include number ofsubjects (n), mean, standard deviation (SD), median, minimum, andmaximum. Summaries of change from baseline variables include onlysubjects who had both a baseline value and corresponding value at thetimepoint of interest. Descriptive statistics for categorical datainclude frequency and percentage.

Listings are provided for all collected study data.

The disposition of all subjects randomized in this study are summarizedby treatment group and completion/discontinuation status. Subjects whodiscontinue the study prematurely are summarized by treatment group andreason for discontinuation. The number of subjects in each analysis setwas summarized by treatment group.

All subject data is reviewed for the occurrence of protocol deviations.Prior to database lock, all protocol deviations are reviewed andclassified with respect to the potential to influence experimentaloutcomes. Protocol deviations are summarized by treatment group.

Demographic and other baseline characteristics are summarized bytreatment group using descriptive statistics.

All prior and concomitant medications are coded using the WHO DrugDictionary. Prior and concomitant medications are summarized bytreatment group using descriptive statistics.

Exposure to investigational product is summarized by treatment groupusing descriptive statistics.

Safety analyses are conducted using data from the Safety Analysis Set.Safety variables included TEAEs, clinical laboratory values, vitalsigns, and ECG results. No formal inferential analyses are conducted forany safety variables, unless otherwise noted.

Adverse event verbatim terms are coded using the Medical Dictionary forRegulatory Activities (MedDRA). The overall incidence of subjects havingat least one AE are summarized by treatment group. The incidence ofTEAEs is summarized by treatment group, system organ class (SOC) andpreferred term (PT). Each subject is counted only once per SOC andpreferred term. An AE was considered treatment-emergent if it occurredafter the first dose of investigational product and within 30 days aftera subject's last dose of investigational product.

For all continuous laboratory test variables, descriptive statistics forall reported values and change from baseline values are summarized bytreatment group and time point.

For all continuous vital sign and ECG variables, descriptive statisticsfor all reported values and change from baseline values are summarizedby treatment group and time point. In addition, the frequency andpercentage of subjects with abnormal ECG findings are summarized.

The proportion of subjects alive and free of respiratory failure with90% confidence interval is now presented. In addition, the proportion ofsubjects alive and free of respiratory failure in the anti-LIGHTmonoclonal antibody group has been compared to that in the placebo groupusing a Chi-square test or similar methods. Other dichotomous efficacyvariables are analyzed similarly. All efficacy variables have beensummarized using descriptive statistics.

For all PK variables, descriptive statistics are presented by collectiontimepoint (where applicable) using the PK Analysis Set. Descriptivestatistics for plasma concentrations include n, number of subjects withconcentrations below the level of quantification (BLQ), mean, SD,coefficient of variation, median, minimum, and maximum. For descriptivesummaries, plasma concentrations reported as BLQ are be set to zero.

For all PD variables, descriptive statistics are presented by treatmentgroup and time point.

For all immunogenicity variables, descriptive statistics are presentedby treatment group and time point.

Example 3

Presented herein in Example 3 are certain results of the study conductedaccording to the methods as described in Example 2.

The screening involved identifying patients with COVID-19 associatedpneumonia and mild to moderate ARDs. Then, patients meeting thesecriteria (n=83) were randomized 1:1 to receive standard of caretreatment plus the anti-LIGHT monoclonal antibody or standard of caretreatment plus placebo. One out of 83 patients withdrew informedconsent. A total of 82 patients were administered either standard ofcare treatment plus the anti-LIGHT monoclonal antibody or standard ofcare treatment plus placebo. A full analysis for secondary endpoints andsafety was completed for these patients. An ITT analysis for primaryendpoint was completed in a group of 62 of the patients who were free ofhigh flow 02 or positive pressure 02 with 28-day follow-up.

Greater than 90% of patients in the study received corticosteroids andgreater than 60% received remdesivir. Patient demographics were asfollows in Table 7:

TABLE 7 Patient Demographics Anti-LIGHT monoclonal antibody PlaceboCharacteristic (n = 41) (n = 42) Age, years Mean (SD) 59.2 (14.5) 58.1(14.2) Age Group <60 years (n, %) 20 (48.8%) 21 (50.0%) ≥60 years (n, %)21 (51.2%) 21 (50.0%) Gender Male 25 (61%) 32 (76.2%) Female 16 (39%) 10(23.8%) Race White 31 (75.1%) 37 (88.1%) Black or African American 7(17.1%) 3 (7.1%) Asian 2 (4.9%) 0 (0%) Other 1 (2.4%) 2 (4.8%) FreeLIGHT Level at Baseline Mean (range) pg/mL 348 (63-1050) 273 (37-843)Concomitant Medication Use at Baseline* Systemic corticosteroids 38(95.0%) 37 (88.1%) Remdesivir 26 (65.0%) 28 (66.7%) *Calculated frompatients dosed (n = 40 for the anti-LIGHT monoclonal antibody, n = 42for placebo).

The study demonstrated robust improvement in the primary endpoint(proportion of patients alive and free of respiratory failure over the28-day study period) compared to placebo in COVID-19 patients with ARDStreated with a single dose of the anti-LIGHT monoclonal antibody at 16mg/kg (max 1,200 mg) (n=62), as shown in FIG. 10 . A prespecifiedsubpopulation of patients greater than or equal to 60 years of ageshowed similar improvement in the primary endpoint (n=34); efficacy washighest in this subpopulation, the population most vulnerable to severecomplications and death with COVID-19 infection. Patients treated withthe anti-LIGHT monoclonal antibody in this subpopulation also had ashorter hospital stay compared with placebo treated patients.

Further, due to the protocol allowing patients to receive high flowoxygen prior to randomization, 62 patients were included in theintention-to-treat (ITT) analysis of the primary endpoint. There was anumerical mortality benefit favoring the anti-LIGHT monoclonal antibodywith 4 patients dying on active drug and 9 on placebo as of the date ofdata retrieval.

A single dose of anti-LIGHT monoclonal antibody reduced mortality be˜50% in the study, observed at both the 28-day and 60-day timepoints.28-day mortality was substantially reduced in patients treated with theanti-LIGHT monoclonal antibody (3 patients) placebo (6 patients). 28-daymortality was 7.7% in patients treated with the anti-LIGHT monoclonalantibody and 14.3% in patients treated with placebo. 60-day mortalitywas 10.8% in patients treated with the anti-LIGHT monoclonal antibodyand 22.5% in patients treated with placebo.

>90% of patients in the study received corticosteroids and >65% receivedremdesivir. Thus the anti-LIGHT monoclonal antibody showed statisticallysignificant efficacy on top of corticosteroids and standard of care inCOVID-19 ARDS.

The anti-LIGHT monoclonal antibody dramatically and rapidly reducedserum free-LIGHT levels in these patients (about 85% reduction in freeLIGHT in one day). See FIG. 9 , where square boxes are subjects treatedwith placebo, circles are subjects treated with the anti-LIGHTmonoclonal antibody. Table 8 shows LIGHT levels over time for patientsin the study who received placebo (n, mean, SD, min and max), and Table9 shows LIGHT levels over time for patients in the study who receivedthe anti-LIGHT monoclonal antibody (n, mean, SD, min and max).

TABLE 8 LIGHT Levels for Patients Treated With Placebo Day n Mean SD MinMax 1 39 276.025641 204.181741 37 843 2 38 303.421053 195.424366 73 7605 22 349.045455 187.021129 130 897 8 12 427.25 285.03688 98 932 9 13459.307692 346.335672 86 1240 14 8 685.625 1131.91884 95 3420 28 19408.157895 309.366425 71 1160

TABLE 9 LIGHT Levels for Patients Treated With Anti-LIGHT Mab Day n MeanSD Min Max 1 40 329.425 241.33271931 22 1050 2 39 51.72820512859.143349267 5.7 360 5 31 42.64516129 51.900898121 5 251 8 15 57.260.74678122 13 239 9 10 51.9 34.326698388 12 123 14 4 59.75 23.51418011927 80 28 25 24.988 13.674255617 5.3 63

Table 10 shows IL-6 levels over time for patients in the study whoreceived placebo (n, mean, SD, min and max), and Table 11 shows IL-6levels over time for patients in the study who received the anti-LIGHTmonoclonal antibody (n, mean, SD, min and max).

TABLE 10 IL-6 Levels for Patients Treated With Placebo Day n Mean SD MinMax 1 39 18.015384615 40.223090308 2.85 190 2 38 10.07105263215.096422664 2.85 64 5 22 17.904545455 26.750664919 2.85 110 8 1228.508333333 36.723827002 2.85 100 9 13 59.473076923 152.07703313 2.85557 14 8 35.8 42.056042542 2.85 106 28 19 26.507894737 54.277297953 2.85187

TABLE 11 IL-6 Levels for Patients Treated With Anti-LIGHT Mab Day n MeanSD Min Max 1 40 11.855 19.83956097 2.85 117 2 39 9.34102564117.152109245 2.85 84 5 31 12.191935484 21.311671672 2.85 93 8 1511.123333333 10.204968444 2.85 36 9 10 12.84 9.7115852923 2.85 29 14 4358.225 707.85297026 2.85 1420 28 25 4.992 4.6357011336 2.85 24

The anti-LIGHT monoclonal antibody was safe and well tolerated with noappreciable differences in immunosuppression or other SAE between theanti-LIGHT monoclonal antibody and placebo. Specifically, the anti-LIGHTmonoclonal antibody was well-tolerated at a single dose of 16 mg/kg; noserious adverse events were attributable to the anti-LIGHT monoclonalantibody; the majority of AEs were judged to be mild or moderate; andthere was no evidence of increased infections or adverse events relatedto immunosuppression (see Table 12 below).

TABLE 12 Adverse Events Anti-LIGHT monoclonal antibody Placebo (n = 40)(n = 42) Subjects with ≥1 AE (%) 16 (40%) 21 (50%) Subjects with ≥1Drug-related AE 8 (20%) 6 (14.3%) AEs > 5% Leukocytosis 6 (15%) 4 (9.5%)Anemia 4 (10%) 3 (7.1%) Hepatic enzyme increase 4 (10%) 2 (4.8%) Acutekidney injury 3 (7.5%) 2 (4.8%) Respiratory failure 3 (7.5%) 3 (7.1%)

Example 4 Free Light Detection Assay Capture of Free Light withCandidate Pair

Simoa ultra-high sensitive assay (Myriad RBM) was used to detect andmeasure free LIGHT with high sensitivity, using Quanterix's fullyautomated immunoassay platform: Simoa HD-1 Analyzer and single moleculearray (Simoa) technology. All incubations take place at room temperatureinside the Simoa HD-1 analyzer. Capture antibody conjugated paramagneticbeads were incubated with standards, samples or controls andbiotinylated detection antibodies. The beads were then washed andincubated with streptavidin-ß-galactosidase (SßG). After the final wash,the beads were loaded into the Simoa Disc with enzyme substrate,resorufin ß-galactopyranoside (RGP). The fluorescence signals arecompared to the standard curve and the quantity of LIGHT Free isdetermined for each sample. After screening anti-LIGHT antibodies inpairs for sandwich immunoassay, assays with one candidate pair (captureantibody: Enzo ALX-804-841-C100, detection antibody: ProSci RF16062;specific epitopes are not detailed for these antibodies) were performedto test for linearity and specificity.

Linearity is a measure of the analytes acceptable sample dilution, andis measured by the ability to obtain results proportional to the analyteconcentration in sample when serially diluted. It showed that samples asdiluted as 2-fold, 4-fold, 5-old, 10-fold, 20-fold, 40-fold, 80-foldmaintained linearity. Linearity conducted at 1:10, 1:20, and 1:40 isshown in FIG. 1 . Linearity conducted at 1:10, 1:20, 1:40, 1:80 is shownin FIG. 2 .

Free LIGHT Detecting Antibody Pair, DCR3 Interference

DcR3 interference was tested on the candidate pair EnzoALX-804-841-C100-ProSci RF16062 (capture-detection). A diluent usingcontaining free LIGHT was used, rather than native free LIGHT in a serumor plasma sample. DcR3 spiked concentration was 10,000 ng/ml and 11additional lower concentrations. Signal inhibition value was calculatedas a signal reduction (mean fluorescence intensity (MFI)) for theEnzo/ProSci pair as shown in FIG. 3 . DcR3 exhibits 96.3% of signalinhibition (=(9938-361)/9938) for the candidate pair. This demonstratedthat the epitope of free LIGHT for the examined pair overlaps with theepitope of free LIGHT for DcR3. It thus shows that the candidate pairbinds free LIGHT.

Free LIGHT Detecting Antibody Pair, DcR3 Interference Comparison

Spike and recovery experiment was performed to assess DcR3 (10 μg/mL)interference on the candidate pair Enzo ALX-804-841-C100-ProSci RF16062(capture-detection). Serum and plasma samples containing native freeLIGHT were incubated with (spiked) and without (unspiked) 150 pg/mL offree LIGHT (LIGHT standard recombinant antigen). Said spiked andunspiked samples were then incubated with DcR3.

The % recovery signal was calculated compared to the control with nointerferent based on the MFI data as shown in FIG. 4A. The % recoverysignal with the interference is divided by the signal of the control.The signal inhibition value was calculated for unspiked serum 1 (whichhad the most significant reduction). In the set shown in FIG. 4A, 78%represents the reduction in signal, which is related (100%-% recovery).That is, serum 1's DcR3 recovery is 22%, representing a 78% reduction insignal. In addition, in the group of LIGHT (150 pg/ml) spiked samples(lower panel), sample serum 1 demonstrates 92% inhibition (8% recovery).

Another set of data for the same experiment, but using ProSciRF16062-LSBio LS-C133566-100 (capture-detection) is shown in in FIG. 4B.

A graph characterizing DcR3 interference with recovery for eachcandidate pair in native free LIGHT and spiked free LIGHT samples isshown in FIG. 4C. The candidate pair Enzo ALX-804-841-C100-ProSciRF16062 (capture-detection) was identified as a native free LIGHTbinder, since it competes with DcR3 and an anti-LIGHT antibody to anextent of above 60% competition. For this pair, serum was a preferablesample to detect free LIGHT compared to plasma. Sensitivity, althoughnot fully characterized yet, implied that samples dilution can be10-fold, which reduces background. In contrast, the relatively littleDcR3 interference with % recovery signal for candidate pair ProSciRF16062-LSBio LS-C133566-100 (capture-detection) indicates the pair doesnot as effectively bind native free LIGHT, even though both candidatepairs bound to non-native LIGHT standard recombinant antigen to aboutthe same degree.

Assay Validation

The Free LIGHT assay with the free LIGHT binder pair was tested for theassay validation parameters of least detectable dose, lower limit ofquantitation, upper limit of quantitation, dynamic range, precision,spike recovery, linearity, matrix interferences, freeze-thaw stabilityand short-term analyte stability, and met the acceptance criteria forthe above listed parameters and reproducibly quantitates free LIGHTlevels in serum/plasma samples.

Antibody competition was performed to assess if any unspecific bindingoccurred. Serum and Plasma samples were incubated with and without 50μg/mL of capture antibody for 35 minutes at room temperature prior toanalysis on the Quanterix HD-1 instrument. Percent recovery wascalculated as capture antibody treated (competitive) sample vs.non-treated (control) sample concentration. The % Recovery was all 0%,well within the acceptance criterion of ≤20%.

Example 5 Free Light Levels in Crohn's Disease Samples

FIG. 5 shows free LIGHT levels in serum samples from 89 Crohn's Disease(CD) subjects were selected and grouped according to time from illness.89 subjects and 10 healthy controls (gender and age matched) weremeasured using the free-LIGHT assay described herein using the candidateantibody pair. After excluding outliers, 62 samples and 7 controls wereanalyzed. Crohn's Disease subjects showed significantly higher serumfree LIGHT levels (527.93 pg/ml, average in subjects of 0-1 month fromillness) than in healthy controls (40.43 pg/ml; P<0.0021). Free LIGHTserum levels also correlated with the disease programs. This suggeststhat Free LIGHT represents a potential target for the treatment of CDand free LIGHT assay can serve as a companion diagnostic for anti-LIGHTtherapy.

Example 6

This study is an escalating dose, open-label, signal-finding study toevaluate the safety, tolerability, and short-term efficacy of ananti-LIGHT monoclonal antibody in adults with moderate to severe activeCrohn's Disease who have failed prior treatment with an anti-TNFα agent,with and without loss of function mutations in decoy receptor 3 (DcR3).The anti-LIGHT antibody has a VH of SEQ ID NO. 8, and a VL of SEQ ID NO:9.

Example 6.1—Study Objectives and Endpoints

The primary objective of this study is to evaluate the safety andtolerability of the anti-LIGHT monoclonal antibody administered by SQinjection to adults with moderate to severe, active CD who have failedprior treatment with an anti-tumor necrosis factor alpha (anti-TNFα)agent.

The secondary objectives of this study are to: estimate plasmaconcentrations of the anti-LIGHT monoclonal antibody administered by SQinjection to adults with moderate to severe, active CD; and to evaluateresponse to treatment with the anti-LIGHT monoclonal antibodyadministered by SQ injection to adults with moderate to severe, activeCD.

Example 6.2—Study Design

This is a Phase 1b, multi-center, open-label, dose-escalation,signal-finding study to evaluate the safety, tolerability, PK andshort-term efficacy of the anti-LIGHT monoclonal antibody in adults withmoderate to severe, active CD who have previously failed anti-TNFαtreatment.

Four subjects with Crohn's Disease who satisfy all eligibility criteriaare enrolled in each of 2 dose cohorts. The first cohort receives theanti-LIGHT monoclonal antibody 1.0 mg/kg SQ every 14 (q14) days.

Dose escalation proceeds after completion of the first cohort based onreview of cumulative safety, tolerability, pharmacokinetic, and efficacydata by Data Monitoring Committee and after a decision is made toprogress to the second cohort. The estimated dose escalation for thesecond cohort is 3.0 mg/kg SQ q14 days, if permitted by safety datareview.

Each subject's participation includes a screening period, which ifrequired includes a 12-week wash-out period for subjects receivingbiologic treatment or who have received biologic treatment within 12weeks of the Screening Visit. For subjects requiring wash-out, there isoptionally a 1- to 14-day time period between the screening visit andthe start of the wash-out period, as necessary. With the exception ofsubjects requiring wash-out of the biologic certolizumab pegol (Cimzia),only those subjects without detectable biologic levels after 8 weeks ofwash-out are allowed to enter the study after confirmation ofundetectable levels; all other subjects (including those receivingcertolizumab pegol [Cimzia]) are required to complete a full 12-weekwash-out period. The wash-out period includes the time period from thelast dose received prior to the Screening Visit. Subjects not requiringa biologic wash-out period are allowed to enter the study after reviewand confirmation of eligibility at screening. Screening is followed byan 8-week, open-label treatment period, and a safety follow-up visitapproximately 4 weeks after the last dose. The maximum study duration is26 weeks.

Study visits occur at screening and on Days 0, 7, 14, 21, 28, 35, 42, 49and 56. The safety follow-up visit occurs on Day 84. The Schedule ofAssessments is shown in Table 13.

TABLE 13 Schedule of Assessments Screening Period Open-Label TreatmentPeriod Visit for Visit 2^(c) Testing Wash- Dosing Visit 1 InitialPrevious out Pre- and Post- Week −14 Phone Biologics Phone dose doseVisit 3 Assessment or Procedure to −12 Contact^(a) (if required)Contact^(b) Day 0 Day 0 Day 7 Informed consent X Inclusion/exclusion X XX criteria review Testing for Previous  X^(q) Biologies Genotyping forDcR3 X genes Demographics/medical X history^(d) Physical examination, XX incl. weight^(e) Vital signs (BP, pulse, X X X RR, T)^(f) TBtesting^(g) X 12-lead ECG X Clinical laboratory X X assessments^(h) PKblood sampling^(i) X X Blood draws for X exploratory analyses^(j)Anti-drug antibody X assessment Urinalysis X X Pregnancy testing(females X X of childbearing potential)^(k) Urine drug screen^(l) XStool sample^(m) X CDAI X X IBD-Q X X Endoscopy with biopsy and Xhistology^(n) Adverse event monitoring^(o) X X X X X X X Concomitantmedications X X X X X X X Provide/reconfirm access X X X to subjectdiary (US sites) Provide diary (ex-US sites) X X X Subject diaryreviewed for X X completeness Assess individual subject X X stoppingcriteria Anti-LIGHT monoclonal X antibody administration (on-site)^(p)Safety Follow-up Open-Label Treatment Period Visit 11 Visit Day 84Assessment 10/ET or 28 or Visit 4 Visit 5 Visit 6 Visit 7 Visit 8 Visit9 Visit days after Procedure Day 14 Day 21 Day 28 Day 35 Day 42 Day 49Day 56 ET visit Informed consent Inclusion/exclusion criteria reviewTesting for Previous Biologies Genotyping for DcR3 genesDemographics/medical history^(d) Physical examination, X X X X incl.weight^(e) Vital signs (BP, pulse, X X X X RR, T)^(f) TB testing^(g)12-lead ECG X X Clinical laboratory X X X X assessments^(h) PK bloodsampling^(i) X X X X X X X X Blood draws for X X exploratoryanalyses^(j) Anti-drug antibody X X X X assessment Urinalysis X XPregnancy testing (females X X of childbearing potential)^(k) Urine drugscreen^(l) Stool sample^(m) X X CDAI X X X X IBD-Q X Endoscopy withbiopsy and X histology^(n) Adverse event monitoring^(o) X X X X X X X XConcomitant medications X X X X X X X X Provide/reconfirm access X X X XX X X to subject diary (US sites) Provide diary (ex-US sites) X X X X XX X Subject diary reviewed for X X X X X X X completeness Assessindividual subject X X X X X X X stopping criteria Anti-LIGHT monoclonalX X X antibody administration (on-site)^(p) ^(a)Telephone visit isconducted for confirmation of eligibility and, as needed, initiateswash-out of current anti-TNFα treatment. If the subject is not receivingbiologic treatment, they begin completing the subject diary and havevisit 2 scheduled >7 days from the initial phone contact. The diary iscompleted up to the time of the visit. ^(b)Telephone visit is conductedfor confirmation whether additional wash-out time is required followingreceipt and review of the laboratory results. If the concentration levelis not detectable, the subject begins completing the subject diary andhas visit 2 scheduled >7 days from the washout phone contact. If theconcentration level is detectable, the subject completes the full 12week wash-out. Upon completion of the washout period, the subject beginscompleting the diary and has visit 2 scheduled >7 days from the end ofthe washout. The diary is completed up to the time of the visit.^(c)After Day 0 (Visit 2), visits at which the anti-LIGHT monoclonalantibody is administered occur every 14 ± 3 days. These visits arescheduled relative to Day 0 (Visit 2), which is the baseline visit.^(d)Medical history includes details regarding all CD-related surgicalprocedures and hospitalizations. ^(e)A complete physical examination isperformed at Visits 1, 2, 6, and 10/ET. Brief physical examinations aredone at Visits 4 and 8. Height is measured at V2 only. ^(f)Vitals aretaken pre-and post-dose on Visit 2, 4, 6 and 8. Post-dose vitals aretaken at least 60 minutes post-dose, immediately prior to discharge.^(g)TB testing includes QuantiFERON-TB Gold (QFT) blood testing or PPDskin testing. If the subject's PPD tine skin test is ≥5 mm, a chestx-ray is also be employed for TB assessment. ^(h)Clinical laboratoryassessments at Visits 1, 6, and 10/ET include: CBC with differential,hematology, serum albumin, CRP, liver panel, GGT. Anti-drug antibodiesare measured at Visits 2, 6 and 10. Clinical laboratory assessments atVisits 2, 4, and, 8 include hematology and serum albumin. ^(i)Pre-dosePK blood samples are obtained within 60 minutes prior to dose at Visits2, 4, 6 and 8. ^(j)Exploratory analyses examined LIGHT, cytokines, RNAsequencing, and flow cytometry. ^(k)Serum β-hCG is conducted at Visit 1and Urine β-hCG tests are conducted on Visits 2, 6 and 10. ^(l)Screeningfor amphetamines, barbiturates, benzodiazepines, cocaine, opiates,phencyclidine, cannabinoids, propoxyphene, and methadone as warranted.^(m)Stool samples are obtained at Visits 1, 6 and 10/ET. C. difficiletest is performed only at Visit 1. Fecal calprotectin is measured at allspecified visits. ^(n)Histological confirmation of disease is performedonly at Visit 1. Documentation is provided to site for confirmation.Exploratory biomarker histology is completedfor samples provided tostudy central laboratory provider. ^(o)Adverse event monitoring beginsat the time informed consent is signed. ^(p)Subjects are required toremain in the clinic for at least 60 minutes after the anti-LIGHTmonoclonal antibody administration for adverse event monitoring.^(q)Subjects requiring wash-out have a blood test performed at 8 weeksafter the start of the wash-out period to confirm the previous biologictherapy is undetectable. If there is no previous biologic therapydetected, subjects continue onto Visit 2 and are not required tocomplete the full 12-week wash-out. If there are detectable levels ofthe previous biologic therapy, then subjects complete the full 12-weekwash-out before proceeding to Visit 2. For subjects receivingcertolizumab pego1 (Cimzia) a 12- week wash-out is required. BP = bloodpressure; CBC = complete blood count; CDAI = Crohn's Disease ActivityIndex; CRP = C-reactive protein; DcR3 = decoy receptor 3; GGT =gamma-glutamyl transferase; ECG = electrocardiogram; ET = earlytermination; hCG = human chorionic gonadotropin; LIGHT =Lymphotoxin-like, exhibits Inducible expression, and competes withHerpes Virus Glycoprotein D for Herpesvirus Entry Mediator, a receptorexpressed by T lymphocytes; PK = pharmacokinetic; PPD = purified proteinderivative; RR = respiration rate; T = temperature; TB = tuberculosis;TNFα = tumor necrosis factor alphaStudy Periods

The study includes a screening period, an open-label treatment period,and a safety follow-up visit. The 1.0 mg/kg dose cohort completes thestudy periods described below. Following a review of the safety data, adecision is made.

Screening Period

During the screening period (Week −14 to Day 0), all subjects areevaluated for their eligibility to participate in the study. At Visit 1,subjects sign the informed consent form before any study-relatedprocedures or evaluations are conducted.

Demographic information is obtained along with medical history,including CD diagnostic information, CD-related procedures/surgeries andmedication use history. Any existing conditions reported at thescreening visit are recorded in the electronic case report form (eCRF);current medications are also recorded on the eCRF. Prior CD therapies(lifetime recall) are recorded in the eCRF with dates reflecting prioruse. All safety assessments are conducted, including physicalexaminations (with weight measurements), vital signs (blood pressure,pulse, respiration rate and temperature), 12-lead ECGs, clinicallaboratory tests, stool sample and urinalysis. A urine drug screeningtest (as warranted) and tuberculosis (TB) testing defined as either apurified protein derivative (PPD) skin reaction test or QuantiFERON-TBGold (QFT) blood test, and as required by protocol, chest x-ray, areadministered to all subjects. A serum β-human chorionic gonadotropin(β-hCG) test is to be administered to females of childbearing potential.The CDAI and the Inflammatory Bowel Disease Questionnaire (IBD-Q) areadministered. Subjects undergo an endoscopy with biopsy and histologyduring the screening period. Subjects are provided access to a studydiary (electronic or hard copy) to record their daily assessment ofwell-being, abdominal pain and stool frequency including loose andwatery stools. The diary is completed for a minimum of 7 daysimmediately prior to initiation of open-label treatment as described inTable 13. Subjects record frequency of loose or watery stools and assessoverall abdominal pain for each day. Finally, adverse events (AEs) aremonitored.

Subjects who are taking a biologic treatment or who have receivedbiologic treatment within 12 weeks of the Screening Visit but who areotherwise eligible for study participation based on review of allscreening evaluations and results begin a wash-out period for thesemedications. With the exception of subjects requiring wash-out of thebiologic certolizumab pegol (Cimzia), a blood test is administered tosubjects undergoing wash-out 8 weeks after their last dose of biologictreatment to ensure that serum levels of any previous biologictreatments are below the level of detection. If it is confirmed that thelevels of previous biologic treatment are undetectable, then the subjectproceeds to Visit 2. If the test results indicate that the previousbiologic treatment is detectable, the subject is required to complete afull 12-week wash-out period before proceeding to Visit 2. For subjectsreceiving the biologic certolizumab pegol (Cimzia) a 12-week wash-out isrequired.

Open-Label Treatment Period

Eligible subjects return to the clinic after the screening period (andwash-out period, if applicable) on Visit 2, which is Day 0 of the8-week, open-label treatment period. Physical examinations includingweight and vital sign assessments, clinical laboratory tests,urinalysis, and the CDAI are performed. A urine β-hCG test isadministered to females of childbearing potential. Any AEs andconcomitant medications are recorded. Blood is drawn for PK, ADA andexploratory analyses (including LIGHT, cytokines, RNA sequencing, andflow cytometry). Subjects record their daily assessment of well-being,abdominal pain and stool frequency including loose and watery stools intheir study diary. Diary data is reviewed for completeness during eachvisit. Subjects are re-evaluated to determine whether they meet allinclusion criteria and do not satisfy any of the exclusion criteria.

Eligible subjects are then enrolled in the study and receive their firstdose of the anti-LIGHT monoclonal antibody in the clinic. Subjectsreceive the anti-LIGHT monoclonal antibody as a SQ injection in theabdomen in a zone of 4 to 10 cm from the umbilicus with the injectionsite rotated with each subsequent dose. The dose of the anti-LIGHTmonoclonal antibody is administered every 14 days (±3 days) for 8 weeks.Subjects are monitored for AEs during the administration of each doseand for 60 minutes after dosing.

Subject enrollment within a cohort is staggered by at least one day, inorder to assess any delayed adverse events.

Subjects return to the clinic every 14 (±3) days (Visits 4, 6, 8, and10) after the first anti-LIGHT monoclonal antibody dose for assessmentof safety, PK, and efficacy and to receive the next dose ofinvestigational product, as shown in Table 13. Additional weekly visitsbetween dosing visits (Visits 3, 5, 7, and 9) occur at which time bloodare drawn for PK analyses, subjects are provided access to a study diary(electronic or hard copy) for diary data collection, and AEs andconcomitant medication use are recorded.

Prior to enrolling subjects in the second dose cohort, all subjects inthe first cohort complete their participation in the study and safetydata from those subjects is reviewed by the Data Monitoring Committee. Arecommendation is provided by the DMC and a decision is made by theSponsor as to whether subjects can be enrolled in the next dose cohort.

Safety Follow-Up Visit

Approximately 28 days after the final dose, subjects have a safetyfollow-up visit. The safety follow-up visit is conducted in the clinicwith the subject. Any AEs that occur in the time since the subject'slast anti-LIGHT monoclonal antibody dose are recorded, along with anyconcomitant medication use.

Safety Data Review

Each investigator revies all available safety data collected at theirsite on a weekly basis, and communicates any safety concerns to thesponsor medical monitor. The sponsor medical monitor reviews allrelevant safety findings with the coordinating principal investigatorand DMC as needed.

Special attention is paid to adverse drug reactions observed in theanti-LIGHT monoclonal antibody preclinical study as well as other commonadverse reactions seen with other biologic treatments which includeinjection site reactions for the anti-LIGHT monoclonal antibody duringpreclinical observation. Special attention is paid to adverse reactionsobserved with other biologic treatments, which include: potential forincreased infection including opportunistic infections (such astuberculosis); hypersensitivity reactions (including anaphylaxis);immunogenicity; malignancy; impaired immunization; and CD exacerbation.

Data Monitoring Committee

An external, independent Data Monitoring Committee (DMC) comprisingphysicians, scientists and a biostatistician review the study data atregular intervals for the duration of the study which includes a meetingafter the completion of each cohort and ad hoc meetings to assessindividual exacerbations of Crohn's Disease meeting Common TerminologyCriteria for Adverse Events (CTCAE) Grade 3 or greater. The DMC's roleis to protect the interests of the subjects in the study and those stillto be entered in the study by reviewing cumulative safety, tolerability,pharmacokinetic and efficacy data. The DMC's meeting schedule may beadjusted based on recommendations made by the DMC, the amount ofincremental safety data, and other practical considerations. The dataprovided to the DMC may not be monitored and is not considered “clean”until the database is locked at the completion of the study.

Possible outcomes of the DMC review can include one of the followingrecommendations: study can continue; study can continue withmodifications; and study is to be terminated.

Data Monitoring Committee recommendations are documented in meetingminutes which include, at a minimum: a list of meeting participants; asummary of data considered during the meeting; and a summary of the DMCrecommendation regarding further dose cohorts, including any concernsraised.

The sponsor is responsible for the decision to continue, modify orterminate the study. A copy of the DMC meeting recommendation andsponsor decision are sent to the study sites upon completion and priorto administration of the next subject dose or initiation of the nextcohort.

Individual Subject Stopping Criteria

The following individual subject stopping criteria are used during thestudy and are assessed starting post-dose at Visit 2. The individualsubject is stopped from the study if the subject develops a CTCAE Grade3 or higher of the following: injection site reactions; opportunisticinfections (i.e., tuberculosis); hypersensitivity reactions (e.g.allergic reactions, anaphylaxis or cytokine release syndrome);malignancy; decreased white blood cell count; decreased neutrophilcount; decreased platelets; colonic or ileal hemorrhage; colonic, ilealor small intestine obstruction; colonic, ileal or small intestineperforation; and colonic, ileal or small intestine stenosis.

A subject is also to be stopped if liver enzymes are: ALT or AST >8×ULN;ALT or AST >5×ULN for more than 2 consecutive weeks or; a single subjectALT or AST >3×ULN with the appearance of fatigue, nausea, vomiting,right upper quadrant pain or tenderness, fever, rash and or eosinophilia(>5%). Note that subjects who exhibit ALT or AST >3×ULN withoutappearance of any of the above symptoms are required to have repeattesting within 48-72 hours to confirm abnormality and determinedirection (increase or decrease) from the original value. A subject isstopped if Hy's Law is detected (i.e. ALT or AST >3×ULN and totalbilirubin >2×ULN and no other reason can be found to explain thecombination of increased AT and TBL, such as viral hepatitis A, B, or C;preexisting or acute liver disease; or another drug capable of causingthe observed injury).

Study Stopping Criteria

The following study stopping criteria are used during the study. Thestudy is stopped if two or more subjects develop the same CTCAE Grade 3or if one subject develops a CTCAE Grade 4 of the following: injectionsite reactions; opportunistic infections (i.e. tuberculosis);hypersensitivity reactions (e.g. allergic reactions, anaphylaxis orcytokine release syndrome); malignancy; decreased white blood cellcount; decrease neutrophil count; or decreased platelets. The study isstopped if two individual subjects develop any of the following livertoxicities: ALT or AST >8×ULN; ALT or AST >5×ULN for more thanconsecutive 2 weeks; or ALT or AST >3×ULN with the appearance offatigue, nausea, vomiting, right upper quadrant pain or tenderness,fever, rash and or eosinophilia (>5%).

The study is stopped if one subject meeting Hy's Law is detected (i.e.ALT or AST>3× ULN and total bilirubin >2× ULN and no other reason can befound to explain the combination of increased AT and TBL, such as viralhepatitis A, B, or C; preexisting or acute liver disease; or anotherdrug capable of causing the observed injury).

Individual reports of exacerbations of Crohn's Disease CTCAE Grade 3 orgreater, such as: colonic or ileal hemorrhage; colonic, ileal or smallintestine obstruction; colonic, ileal or small intestine perforation; orcolonic, ileal or small intestine stenosis are reviewed by the DMC atthe first available date after report to determine if modifying thestudy or stopping the study is recommended.

Study Design Rationale

This is the second study in which the anti-LIGHT monoclonal antibody isadministered to human subjects and the first study in subjects withtreatment-resistant CD. The study is a pilot study using adose-escalation design to characterize the safety and tolerability of 2different doses of the anti-LIGHT monoclonal antibody (1.0 mg/kg and 3.0mg/kg) in the target population. The dose escalation design allows forthe evaluation of safety and tolerability in small numbers of subjectsbefore proceeding to the next dose level. The open-label administrationof the anti-LIGHT monoclonal antibody to all enrolled subjects minimizesthe number of subjects exposed to the study procedures. The inclusion ofPK blood draws and efficacy assessments allowed for preliminaryassessments of plasma levels and efficacy, respectively.

No specific hypotheses are being tested in this pilot study. All dataare to be summarized using descriptive statistics as appropriate.

Number of Subjects

Four subjects are enrolled in each of the 2 planned dose cohorts for amaximum of 8 study subjects. Subjects who withdraw from the studyprematurely prior to a third dose are permitted to be replaced.

Treatment Assignment

The first cohort of subjects are assigned to the 1.0 mg/kg dose of theanti-LIGHT monoclonal antibody. Subjects are assigned to the second dosecohort, after the DMC review of the safety data from the first cohort,and provided study stopping criteria are not met.

Subject Inclusion Criteria

Subjects that meet all of the following inclusion criteria are eligiblefor enrollment in the study:

-   -   1. Subject is able to speak English fluently and provided        written informed consent for this study.    -   2. Subject is male or female, ≥18 to ≤75 years of age.    -   3. Subject has a documented diagnosis of CD via        endoscopy/colonoscopy and histologicalconfirmation.    -   4. Subject has moderate to severe, active CD as evidenced by        Simple Endoscopy Score for Crohn's Disease (SES-CD) score of ≥7        and histological confirmation.    -   5. Subject has failed treatment with an approved therapeutic        dose of an anti-TNFα monoclonal antibody treatment with either        no initial response (primary non-responder) or an initial        response to induction with subsequent lost response (secondary        non-responder) as defined below.    -   6. Subject is permitted to receive concurrent treatment with an        oral corticosteroid, and/orazathioprine or 6-mercaptopurine        (6-MP) or methotrexate (MTX).    -   7. Subject agrees to be genotyped at the DcR3 locus.

A primary non-responder is defined as a subject for whom treatment withinfliximab, adalimumab, or certolizumab pegol produced an inadequateinitial response. Inadequate initial response symptom details occur ≥2weeks after the last dose of induction therapy. The algorithm fordefining inadequate initial response is shown below. Subjectscategorized as primary non-responders meet both parts of the algorithm.Documentation required includes dates and doses of failed inductiontherapy and lack of response details around disease activity recorded bya treating clinician.

The following algorithm is used for inadequate initial response tocurrent or prior therapy with infliximab, adalimumab, or certolizumabpegol. The subject has received induction doses of either infliximab (2or 3 doses of ≥5 mg/kg), adalimumab (dose of 160 mg followed by a doseof ≥80 mg or, dose of 80 mg followed by a dose of ≥40 mg), orcertolizumab pegol (2 or 3 doses of ≥400 mg); and the subject did notinitially respond to these induction doses as documented by the presenceof at least 1 of the following signs or symptoms related to Crohn'sDisease activity: lack of improvement or worsening in stool frequency;lack of improvement or worsening in daily abdominal pain; occurrence,lack of improvement, or worsening of fever associated with Crohn'sDisease; recurring drainage from a previously non-draining fistula ordevelopment of a new draining fistula; lack of improvement or worseningin rectal bleeding; or initiation or increase in antidiarrhealmedication.

A secondary non-responder is defined as a subject for whom treatmentwith infliximab, adalimumab, or certolizumab pegol produced an initialresponse followed by a loss of response. Loss of response detailsoccurs >2 weeks after last dose of maintenance therapy. The algorithmfor defining loss of response is described below. Subjects categorizedas secondary non-responders meet both parts of the algorithm.Documentation required includes dates and doses of induction andmaintenance, initial response and subsequent loss of response includingdetails around disease activity recorded by a treating clinician. Thefollowing is the algorithm for loss of response to prior therapy withinfliximab, adalimumab, or certolizumab pegol. The subject responded toinduction therapy at doses described above and received at least 2maintenance doses of: infliximab (>5 mg/kg), adalimumab (dose >40 mg or,if failed as a pediatric dose of >20 mg), or certolizumab pegol (>400mg); and the subject did not respond to these maintenance doses asdocumented by the presence of at least 1 of the following signs orsymptoms related to Crohn's Disease activity: worsening in stoolfrequency; worsening in daily abdominal pain; occurrence, or worseningof fever associated with Crohn's Disease; recurring drainage from apreviously non-draining fistula or development of a new drainingfistula; worsening in rectal bleeding; or initiation or increase inantidiarrheal medication.

Subject Exclusion Criteria

Subjects who meet any of the following exclusion criteria are noteligible for enrollment in the study:

-   -   1. Subject has a diagnosis of ulcerative colitis (UC) or        indeterminate colitis.    -   2. Subject is unable to tolerate or unwilling to undergo study        procedures including endoscopy and biopsy during the study.    -   3. Subject has signs or symptoms of bowel obstruction with small        bowel imaging supporting obstruction.    -   4. Subject has short bowel syndrome as determined by the        investigator.    -   5. Subject has a current functional colostomy or ileostomy.    -   6. Subject had a surgical bowel resection within the past 6        months prior to screening or is planning any resection during        the study period.    -   7. Clinical suspicion of intra-abdominal abscesses exist, in the        opinion of the investigator.    -   8. Subject has concurrent bowel dysplasia or a history of bowel        dysplasia in the 5 years prior to screening.    -   9. Subject has a known, active and/or positive test for C.        difficile infection.    -   10. Subject has history of or current diagnosis of any cancer        excluding cancers that have been cured by surgical excision        (e.g., non melanoma skin cancers).    -   11. Subject has a history of a lymphoproliferative disorder,        including lymphoma, or signs and symptoms suggestive of        lymphoproliferative disease at any time.    -   12. Subject has history of or active TB infection or positive TB        testing at screening.    -   13. Subject has known concurrent viral hepatitis, or acquired        immune deficiency syndrome (AIDS) or known human        immunodeficiency virus (HIV) infection.    -   14. Subject has been treated with natalizumab (TYSABRI®).    -   15. Subject has not completed his/her primary vaccination series        (particularly hepatitis B, varicella, measles/mumps/rubella)        unless immunity documented with blood titers.    -   16. Subject received any live attenuated vaccine, such as        varicella-zoster, oral polio, or rubella, within 3 months prior        to the baseline visit.    -   17. Subject has any of the following abnormal screening        laboratory test results: clinically significant ECG        abnormalities; aspartate transaminase (AST), alanine        transaminase (ALT) or total bilirubin >ULN; hemoglobin <10 g/dL;        absolute neutrophil count <1500 cell/mm³, or; estimated        glomerular filtration rate <60 mL/min/1.73 m².    -   18. Subject has abnormal vital signs during Screening (Visit 1)        or prior to enrollment at the baseline visit (Visit 2).    -   19. Subject is pregnant or a nursing mother.    -   20. Subject is sexually active and not on effective        contraception.    -   21. Subject has a history of drug abuse that may inhibit        participation in the clinical study.    -   22. Subject has a current or recent history (within 6 months        prior to screening) of significant and severe renal, hepatic,        hematological, gastrointestinal (other than CD or conditions        outlined above), endocrine, pulmonary, cardiac, or neurological        disease.    -   23. Subject has any other clinically significant mental or        physical illness or infection that, in the opinion of the        investigator, might confound the results of the study, pose        additional risk to the subject by their participation, or        prevent or impede the subject from completing the study.    -   24. There is any concern on the part of the investigator        regarding the subject's safety, compliance, or suitability with        respect to his/her participation in the study.        Screen Failure

Subjects who fail inclusion and/or exclusion criteria are allowed berescreened for the study with the prior approval of the sponsor'smedical monitor. In the event of a rescreening, the first screeningvisit is entered into the eCRF as the Screening Visit (Visit 1) and therepeat assessments are entered into the eCRF as an unscheduled visit.

Subject Withdrawal Criteria

All subjects are advised that they are free to withdraw fromparticipation in this study at any time, for any reason, and withoutprejudice. The investigator makes every reasonable attempt to keepsubjects in the study; however, subjects are withdrawn from the study ifthey withdraw consent to participate. For subjects who fail to attendscheduled visits, the investigator attempts to contact them by telephoneor other means to exclude the possibility of an AE being the cause ofwithdrawal. Should that be the cause, the AE is documented, reported,and followed.

The sponsor reserves the right to request the withdrawal of a subjectdue to protocol violations or other reasons.

The investigator also has the right to withdraw subjects from the studyat any time for lack of therapeutic effect that is intolerable orotherwise unacceptable to the subject, for intolerable or unacceptableAEs, intercurrent illness, for meeting the individual subject stoppingcriteria, noncompliance with study procedures, administrative reasons,or in the investigator's opinion, to protect the subject's bestinterests.

If a subject is withdrawn before completing the study, the reason forwithdrawal and the date of discontinuation are recorded on theappropriate eCRF. Whenever possible and reasonable, the evaluations thatare conducted at the completion of the open-label treatment period(i.e., Visit 10) are performed at the time of early termination.

Subjects who withdraw prior to receiving the third dose of study drug,if any, are allowed to be replaced.

All samples are retained according to applicable rules and regulations.Blood and biopsy samples are stored and used for further analysisrelated to this research. Saliva samples are allowed to be used forpurposes related to this research. Samples are given a unique code thatincludes no information that names the subject. Any remaining DNAsamples are stored for future biomarker studies.

Example 6.3—Treatment of Subjects

Description of Investigational Product

Subjects in each dose cohort receive the investigational product as asingle SQ injection in the abdomen in a zone of 4 to 10 cm from theumbilicus with the injection site rotated with each subsequent dose. Thedose of the anti-LIGHT monoclonal antibody is administered on Days 0,14, 28, and 42. After Day 0, injections occur within ±3 days of thescheduled 14-day intervals.

Subjects in the first dose cohort receive the anti-LIGHT monoclonalantibody 1.0 mg/kg for the entire 8-week, open-label treatment period.Data from this cohort are reviewed, after all subjects have completedthe treatment period and its associated assessments and before thesecond cohort is enrolled. It is anticipated that there are a minimum of2 weeks between subject last visit and the review of cumulative safety,tolerability, pharmacokinetic and efficacy data.

Eligible subjects in the second dose cohort receive the anti-LIGHTmonoclonal antibody 3.0 mg/kg for the entire 8-week, open-labeltreatment period. Data from the second cohort are reviewed, after allsubjects have completed the treatment period and its associatedassessments.

Any quality issue noticed with the receipt or use of an investigationalproduct provided by the sponsor (deficiency in condition, packaging,appearance, pertaining documentation, labeling, expiration date, etc.)is promptly communicated to the sponsor, who investigates.

A potential defect in the quality of investigational product provided bythe sponsor may be subject to initiation of a recall procedure by thesponsor. In this case, the investigator is responsible for promptlyaddressing any request made by the sponsor, in order to recall theinvestigational product and eliminate potential hazards.

Permitted and Prohibited Therapies

All prior lifetime CD therapies as well as concomitant medications used(including over-the-counter medications and herbal supplements) arerecorded in the source document and on the appropriate eCRF.

Permitted Therapies

Subjects are permitted to receive concurrent treatment with an oralcorticosteroid, and/or azathioprine, 6-MP or MTX. Subjects are notallowed to have their dose of these medications increased during thestudy. If a dose increase of a concomitant permitted therapy isrequired, the subject is discontinued from the study utilizing the EarlyTermination visit procedures. Concurrent treatment with an oralcorticosteroid, and/or azathioprine, 6-MP or MTX is defined as follows:Oral corticosteroid-Prednisone dose not exceeding 40 mg/day, with astable dose for at least 2 weeks prior to baseline; Azathioprine or6-MP-Azathioprine dose of at least 2 mg/kg/day or 6-MP dose of 1 to 1.5mg/kg/day rounded to the nearest available tablet formulation, or a dosethat is the highest tolerated for the subject, in the opinion of theinvestigator, for at least 8 weeks prior to baseline with a stable dosefor at least 4 weeks prior to baseline; or MTX dose of 25 mg/week duringstudy, either SQ, intramuscularly, or orally, for at least 8 weeks priorto baseline with a stable dose for at least 4 weeks prior to baseline.

Doses of these therapies are permitted to be decreased during the study;however, all doses are within the combinations and dose ranges specifiedabove. These changes are recorded on the concomitant medication eCRF.

For subjects on corticosteroids at the time of study enrollment, weaningduring the study is done according to the following rules:corticosteroid dose of >20 mg and a maximum taper rate per week of 10mg; 10 to <20 mg and a maximum taper rate per week of 5 mg; or <10 mgand a maximum taper rate per week of 2.5 mg. If a deviation from thepermitted concomitant therapy combinations, dose ranges or weaningschedule are necessary, the subject is withdrawn from studyparticipation.

Prohibited Therapies

Concomitant use of biologic treatments during the study is prohibitedincluding use of anakinra (KINERET®, Amgen), abatacept (ORENCIA®,Bristol-Myers Squibb), or tocilizumab (ACTEMRA®, Genentech). A wash-outperiod of up to 12 weeks is required prior to study enrollment (Visit 2)and after confirmation of eligibility from screening proceduresperformed at Visit 1 for all biologic treatments received within 12weeks of the Screening Visit. Prior treatment with natalizumab(TYSABRI®, Biogen) excludes subjects from participation.

Vaccination with live or attenuated virus 3 months prior to screeningand at any time during the study is prohibited.

Contraceptive Methods

Sexually active study participants agree to the use of effectivecontraceptive methods during the study and for the defined period afterthe end of study visit. Approved methods require double barrieraccording to the following algorithm: condom plus intrauterine device orcondom plus hormonal contraceptive. Should any subject be sterilized,the procedure for sterilization is required to have been completed morethan 3 months prior to study screening visit.

A sexually active male participant is required to use one of theabove-described double barrier contraceptive methods during the studyand for 3 months after the end-of-study visit. A sexually active femaleparticipant is required use one of the above-described double barriercontraceptive methods during the study and for 1 month after theend-of-study visit.

Male subjects also agree not to donate sperm for the duration of thestudy and for up to 3 months after the end-of-study visit.

Study participants who are abstinent at the time of study entry agree touse the approved methods described in this section should they becomesexually active during the study.

Treatment Compliance

Treatment with the anti-LIGHT monoclonal antibody is administered bystudy center personnel under direct medical supervision, and anappropriate record is made in the source data by the investigator orhis/her delegate. The investigator or designee records the dosinginformation on the appropriate eCRF page. It is the investigator'sresponsibility to ensure that an accurate record of the administrationof the investigational product is maintained.

Randomization and Blinding

All subjects receive the anti-LIGHT monoclonal antibody in an open-labelmanner.

Treatment after End of Study

After successful enrollment and subsequent completion of or earlytermination from the study, each subject is treated according tostandard clinical practice. In order to support the subject's transitionfrom the clinical study, after care medical expenses such as co-pays andout of pocket medical/treatment associated costs are covered in thetotal amount of $5,000.00 which may be used up to a total 6 monthspost-study exit. Aftercare payments are administered by a third-partyvendor contracted by the Sponsor.

Example 6.4—Investigational Product Materials and ManagementInvestigational Product

The anti-LIGHT monoclonal antibody is the investigational product usedin this study. It is administered in the dosage form 150 mg/mL solution.The unit dose is 1.0 mg/kg or 3.0 mg/kg. The route of administration isSQ injection in the abdomen in a zone of 4 to 10 cm from the umbilicuswith the injection site rotated with each subsequent dose. It is in acolorless to slightly yellowish brown solution. It is manufactured bysanofi-aventis group.

Packaging

All packaging and labeling operations are performed by the sponsor ordesignee according to Good Manufacturing Practice and Good ClinicalPractice (GCP) rules. The investigational product is sent to the studysite by the sponsor or designee. Labeling is in the local language anddependent upon local regulations.

Labeling

The vial and the carton have affixed a label that meet the applicableregulatory requirements.

The investigator saves all unused or partially used medication vials andall empty packaging for final disposition locally (sites in Colombia andIsrael) or by the sponsor (US sites). Syringes used for dosing aretreated as biologic waste and disposed of properly.

Storage

All investigational product is stored between 2° and 8° C. and protectedfrom light. Investigators or other authorized persons (e.g.,pharmacists) are responsible for storing the investigational productprovided by the sponsor in a secure and safe place in accordance withlocal regulations, labeling specifications, institutional policies andprocedures.

Control of storage conditions for the investigational product providedby the sponsor, especially control of temperature (e.g., refrigeratedstorage) and daily temperature monitoring, and information on in-usestability and instructions for handling the investigational product aremanaged according to the rules provided by the sponsor.

Administration

The anti-LIGHT monoclonal antibody is administered by SQ injection inthe abdomen in a zone of 4 to 10 cm from the umbilicus with theinjection site rotated with each subsequent dose.

Accountability

The investigator maintains adequate records showing the receipt,administration, or other disposition of the investigational productincluding the date, lot identifier, dosage, volume administered to eachsubject, and identification of subjects (subject number and initials)who receive the investigational product. The investigator is notpermitted to supply the investigational product to any other location orperson except those named as subinvestigators on the Form FDA 1572,designated study personnel, and subjects in this study. The investigatoris not permitted to dispense the investigational product from any studysites other than those listed on Form FDA 1572. If any of theinvestigational product is not dispensed; is lost, stolen, spilled,unusable; or was received in a damaged container, this information isdocumented and reported to sponsor and appropriate regulatory agencies,as required.

Upon completion of the study, unused investigational product is left inthe original packaging for final disposition locally (sites in Colombiaand Israel) or by the sponsor (US sites). Any partially usedinvestigational product and all empty packaging (e.g., vials) is savedfor final disposition locally (sites in Colombia and Israel) or by thesponsor (US sites) and returned to the sponsor's designee fordestruction.

Handling and Disposal

Investigational product reconciliation is performed at the site by theinvestigator and the monitoring team using treatment log forms anddocumented on the site's investigational product inventory countersignedby the investigator and the monitoring team.

After reconciliation authorization by the sponsor, all used, partiallyused, and unused vials and all original packaging is disposed of locally(sites in Colombia and Israel) or by the sponsor (US sites). Thisprocess is provided to the site by the sponsor's designee.

Example 6.5—Study Procedures and Assessments

Subjects provide written informed consent before any study-relatedprocedures are initiated, including the cessation of prohibitedconcomitant therapy.

For the timing of assessments and procedures throughout the study, referto the Schedule of Assessments (see Table 13). Throughout the study,every reasonable effort is made by study personnel to follow the timingof assessments and procedures in the schedule of events for eachsubject. Visits performed after the Visit 2 baseline visit are scheduledrelative to Visit 2 in order to maintain 56 days of open-label treatmentand administration every 14 days ±3 days). If a subject misses a studyvisit for any reason, the visit is rescheduled as soon as possible. Eachstudy visit window after Visit 2 is ±3 days. Visit procedures areperformed in the order shown however adjustments are allowed to be madeto the order to accommodate site-specific requirements.

Study Periods and Visits: Screening (Visit 1, Week-14 to Day 0)

The subject is screened within 14 weeks before enrollment in the study.The following procedures are performed at screening:

-   -   1. Obtain written informed consent    -   2. Review inclusion/exclusion criteria    -   3. Conduct genotyping for DcR3 genes    -   4. Collect demographic information    -   5. Record medical and medication history    -   6. Perform a complete physical examination, including        measurements of weight    -   7. Collect vital signs, including systolic and diastolic blood        pressures, pulse, respiration rate and temperature    -   8. Perform 12-lead ECG    -   9. Collect blood samples for clinical laboratory tests    -   10. Perform QuantiFERON-TB Gold (QFT) blood test, or PPD        tuberculosis skin test. If the subject's PPD tine skin test is        ≥5 mm, a confirmatory chest x-ray is required.    -   11. Collect blood sample for β-hCG test (for females of        childbearing potential only)    -   12. Collect stool sample for C. difficile and fecal calprotectin        analyses    -   13. Collect urine sample for urinalysis    -   14. Collect urine sample for drug testing as warranted    -   15. Administer the CDAI    -   16. Administer the IBD-Q questionnaire    -   17. Conduct the endoscopy with biopsy    -   18. Assess and record any AEs and concomitant medications    -   19. Provide access to patient diary (electronic or hard copy) to        record daily assessment of abdominal pain, general well-being        and number of stools including loose or watery stool (e.g., 6 or        7 on Bristol stool form scale) and daily abdominal pain severity        (scale of 0 to 10).

Note that stool samples are allowed to be obtained at any time duringthe visit. Subjects who fail to meet clinical laboratory entryrequirements are allowed to be re-tested once as part of the screeningperiod. Extensions of the screening window to accommodate clinicallaboratories re-testing timeframes (excluding endoscopy with biopsy) arepermitted but the total screening time, including endoscopy andwash-out, is not to exceed 16 weeks.

Study Periods and Visits: Eligibility Check/Wash-Out Period (Week-12 toDay 0)

Each subject receives a telephone call no later than Week −12 (ie, 12weeks before the scheduled day of treatment initiation). This callconfirms continued subject eligibility. For subjects not requiring awash-out period after review and confirmation of suitability during thiscall, the subject is instructed to begin recording their daily abdominalpain rating, general well-being assessment and stool frequency includingloose, watery stool frequency in their diary for a minimum of 7 daysimmediately prior to Visit 2. Once completed the subject returns >7 daysfrom the initial phone call for Visit 2. Any changes in concomitantmedication use and any newly occurring AEs since the last evaluation arerecorded.

For subjects requiring a wash-out, if eligible the subject is asked toinitiate wash-out of any current biologic treatment, as necessary. Withthe exception of subjects requiring wash-out of the biologiccertolizumab pegol (Cimzia), a blood test is administered to subjectsundergoing wash-out 8 weeks after their last dose of biologic treatmentto ensure that serum levels of any previous biologic treatments arebelow the level of detection. If it is confirmed that the levels ofprevious biologic treatment are undetectable, then the subject isinstructed to begin recording their daily abdominal pain rating, generalwell-being assessment and stool frequency including loose, watery stoolfrequency in their diary for a minimum of 7 days immediately prior todosing. Once complete the subject returns >7 days from the washout phonecall for their Visit 2. If the test results indicate that the previousbiologic treatment was detectable, the subject is required to complete afull 12-week wash-out period. For subjects receiving the biologiccertolizumab pegol (Cimzia) a 12-week wash-out is required. Uponcompletion of the washout period, the subject begins completing thediary and has Visit 2 scheduled >7 days from the end of the washout. Thediary is completed up to the time of the visit. Any changes inconcomitant medication use and any newly occurring AEs since the lastevaluation are recorded.

If the subject is not eligible, the subject may have any changes inconcomitant medication use and any newly occurring AEs since the lastevaluation recorded and may be removed from screening.

Study Periods and Visits: Open-Label Treatment Period, Baseline Visit(Visit 2, Day 0)

Prior to administration of the first dose of investigational product onDay 0, the following procedures are performed:

-   -   1. Review inclusion/exclusion criteria to confirm continued        eligibility or screen failure status.        -   If the subject is still eligible for the study, the            subsequent procedures are performed. If the subject is not            eligible, any changes in concomitant medication use and any            newly occurring AEs since the last evaluation are recorded            and the subject is removed from screening.    -   2. Perform a complete physical examination, including        measurements of height and weight    -   3. Collect vital signs, including systolic and diastolic blood        pressures, pulse, respiration rate and temperature    -   4. Collect blood sample for clinical laboratory tests    -   5. Collect blood sample for plasma the anti-LIGHT monoclonal        antibody concentration and PK analysis    -   6. Collect blood sample for plasma ADA analysis    -   7. Collect blood sample for exploratory analyses as defined in        Table 13    -   8. Collect urine sample for urinalysis    -   9. Collect urine sample for urine β-hCG test (for females of        childbearing potential only)    -   10. Administer the CDAI and IBD-Q    -   11. Review diary completion    -   12. Assess and record concomitant medications and newly        occurring AEs since the last evaluation

In addition, ongoing AEs from the previous visit are assessed and anychange in their status is recorded.

After completing the assessments, subjects receive the anti-LIGHTmonoclonal antibody by SQ injection in the abdomen in a zone of 4 to 10cm from the umbilicus with the injection site rotated with eachsubsequent dose. Vital sign assessments are conducted within 60 minutespost-dosing.

Subjects remain in the clinic for 60 minutes after the investigationalproduct has been administered. During this time, the following detailsare recorded:

-   -   1. Any AEs occurring after dosing    -   2. Any concomitant medications administered after dosing    -   3. Assess subject response and available results against        individual stopping rules criteria.

After completion of the post-dosing assessments, the subject ispermitted to leave the clinic.

Study Periods and Visits: Open-Label Treatment Period, Visit 3 (Day 7[±3 Days])

Subjects return to the clinic on Day 7 (±3 days), at which time thefollowing procedures are performed:

-   -   1. Collect blood samples for plasma anti-LIGHT monoclonal        antibody concentration and PK analyses    -   2. Assess and record any AEs and concomitant medications    -   3. Review diary completion    -   4. Assess subject response and available results against        individual stopping rules criteria. In addition, any ongoing AEs        from the previous visit are assessed and any change in their        status was to be recorded.        Study Periods and Visits: Visit 4 (Day 14 [±3 Days])

Subjects return to the clinic on Day 14 (±3 days), at which time thefollowing procedures are performed prior to dosing:

-   -   1. Perform a brief physical examination, including measurement        of weight    -   2. Collect vital signs, including systolic and diastolic blood        pressures, pulse, respiration rate and temperature    -   3. Collect blood sample for clinical laboratory tests    -   4. Collect blood samples for plasma anti-LIGHT monoclonal        antibody concentration, PK and ADA analyses    -   5. Administer the CDAI    -   6. Review diary completion    -   7. Assess and record concomitant medication use and any newly        occurring AEs since the last evaluation

In addition, ongoing AEs from the previous visit are assessed and anychange in their status is recorded.

After completing these assessments, subjects receive the anti-LIGHTmonoclonal antibody by SQ injection in the abdomen in a zone of 4 to 10cm from the umbilicus with the injection site rotated with eachsubsequent dose. Vital sign assessments are conducted within 60 minutespost-dosing.

Subjects remain in the clinic for 60 minutes after the investigationalproduct has been administered. During this time, the following detailsare recorded:

-   -   1. Any AEs occurring after dosing    -   2. Any concomitant medications administered after dosing.

After completion of the post-dosing assessments, the subject ispermitted to leave the clinic.

Study Periods and Visits: Visit 5 (Day 21 [±3 Days])

Subjects return to the clinic on Day 21 (±3 days), at which time thefollowing procedures are performed:

-   -   1. Collect blood sample for plasma anti-LIGHT monoclonal        antibody concentration and PK analysis    -   2. Assess and record any AEs and concomitant medications    -   3. Review diary completion    -   4. Assess subject response and available results against        individual stopping rules criteria.

In addition, any ongoing AEs from the previous visit are assessed andany change in their status is recorded.

Study Periods and Visits: Visit 6 (Day 28 [±3 Days])

Subjects return to the clinic on Day 28 (±3 days), at which time thefollowing procedures are performed prior to dosing:

-   -   1. Perform a complete physical examination, including vital sign        assessment and measurement weight    -   2. Collect vital signs, including systolic and diastolic blood        pressures, pulse, respiration rate and temperature    -   3. Perform 12-lead ECG    -   4. Collect blood sample for clinical laboratory tests    -   5. Collect blood samples for plasma anti-LIGHT monoclonal        antibody concentration, PK and ADA analyses    -   6. Collect blood sample for exploratory analyses as defined in        Table 13    -   7. Collect stool for fecal calprotectin analysis    -   8. Collect urine sample for urinalysis    -   9. Collect urine sample for urine β-hCG test (for females of        childbearing potential only)    -   10. Administer the CDAI    -   11. Review diary completion    -   12. Assess and record concomitant medication use and any newly        occurring AEs since the last evaluation

In addition, any ongoing AEs from the previous visit are assessed andany change in their status is recorded.

After completing these assessments, subjects receive the anti-LIGHTmonoclonal antibody by SQ injection in the abdomen in a zone of 4 to 10cm from the umbilicus with the injection site rotated with eachsubsequent dose. Vital sign assessments are conducted within 60 minutespost-dosing.

Subjects remain in the clinic for 60 minutes after the investigationalproduct has been administered. During this time, the following detailsare recorded:

-   -   1. Any AEs occurring after dosing    -   2. Any concomitant medications administered after dosing. NOTE:        Stool samples may be obtained at any time during the visit    -   3. Assess subject response and available results against        individual stopping rules criteria.

After completion of the post-dosing assessments, the subject ispermitted to leave the clinic.

Study Periods and Visits: Visit 7 (Day 35 [±3 Days])

Subjects return to the clinic on Day 35 (±3 days), at which time thefollowing procedures are performed:

-   -   1. Collect blood sample for pharmacokinetic analysis    -   2. Assess and record any AEs and concomitant medications    -   3. Review diary completion    -   4. Assess subject response and available results against        individual stopping rules criteria.

In addition, any ongoing AEs from the previous visit are assessed andany change in their status is recorded.

Study Periods and Visits: Visit 8 (Day 42 [±3 Days])

Subjects return to the clinic on Day 42 (±3 days), at which time thefollowing procedures are performed prior to dosing:

-   -   1. Perform a brief physical examination, including measurement        weight    -   2. Collect vital signs, including systolic and diastolic blood        pressures, pulse, respiration rate and temperature    -   3. Collect blood sample for clinical laboratory tests    -   4. Collect blood sample for plasma anti-LIGHT monoclonal        antibody concentration and PK analysis    -   5. Administer the CDAI    -   6. Review diary completion    -   7. Assess and record any AEs and concomitant medications

In addition, any ongoing AEs from the previous visit are assessed andany change in their status is recorded.

After completing these assessments, subjects receive the anti-LIGHTmonoclonal antibody by SQ injection in the abdomen in a zone of 4 to 10cm from the umbilicus with the injection site rotated with eachsubsequent dose. Vital sign assessments are conducted within 60 minutespost-dosing.

Subjects remain in the clinic for 60 minutes after the investigationalproduct is administered. During this time, the following details arerecorded:

-   -   1. Any AEs occurring after dosing    -   2. Any concomitant medications administered after dosing.    -   3. Assess subject response and available results against        individual stopping rules criteria.

After completion of the post-dosing assessments, the subject ispermitted to leave the clinic.

Study Periods and Visits: Visit 9 (Day 42 [±3 Days])

Subjects return to the clinic on Day 49 (±3 days), at which time thefollowing procedures are performed:

-   -   1. Collect blood sample for plasma anti-LIGHT monoclonal        antibody concentration and PK analysis    -   2. Assess and record any AEs and concomitant medications    -   3. Review diary completion

In addition, any ongoing AEs from the previous visit are assessed andany change in their status is recorded

Study Periods and Visits: Visit 10 (Day 56 [±3 Days])

Subjects return to the clinic on Day 56 (±3 days), at which time thefollowing procedures are performed:

-   -   1. Perform a complete physical examination, including        measurement of weight    -   2. Collect vital signs, including systolic and diastolic blood        pressures, pulse, respiration rate and temperature    -   3. Perform 12-lead ECG    -   4. Collect blood sample for clinical laboratory tests    -   5. Collect blood samples for plasma anti-LIGHT monoclonal        antibody concentration, PK and ADA analysis    -   6. Collect blood sample for exploratory analyses as defined in        Table 13    -   7. Collect stool for fecal calprotectin analysis    -   8. Collect urine sample for urinalysis    -   9. Collect urine sample for urine β-hCG test (for females of        childbearing potential only)    -   10. Administer the CDAI    -   11. Review diary completion    -   12. Administer IBD-Q questionnaire    -   13. Conduct the endoscopy and biopsy    -   14. Assess and record any AEs and concomitant medications    -   15. Assess subject response and available results against        individual stopping rules criteria.

In addition, any ongoing AEs from the previous visit are assessed andany change in their status is recorded.

Note that stool samples are allowed to be obtained at any time duringthe visit.

Early Termination

If a subject is withdrawn from the study for any reason, every effort ismade to conduct all Day 56 (Visit 10) procedures and assessments.

Study Periods and Visits: Safety Follow-Up, Visit 11 (Day 84 [±3 Days])

Subjects return to clinic 28 days (±3 days) after Visit 10 or the earlytermination visit for a safety follow-up visit, as appropriate. Subjectshave blood collected for plasma anti-LIGHT monoclonal antibodyconcentration, PK and ADA analyses. Any concomitant medications andnewly occurring AEs since the last visit are recorded. In addition,ongoing AEs from the previous visit are assessed and any change in theirstatus is recorded.

Study Duration

The overall study duration is approximately 26 weeks (including up to 14weeks of screening [inclusive of wash-out of up to 12 weeks, ifrequired], 56 days of open-label treatment, and a follow-up visitapproximately 28 days after the final dose of investigational product).

The planned sequence and maximum duration of the study periods is asfollows:

-   -   1. Screening period: approximately 14 weeks    -   2. Wash-out period, if applicable: Up to 12 weeks from the        subject's last dose of biological treatment    -   3. Open-label treatment period: 56 days (beginning on Day 0 with        doses administered q14 days)    -   4. Follow-up: 28 days after the last dose of investigational        product. The maximum study duration for each subject is        approximately 26 weeks.

The maximum treatment duration for each subject is approximately 56days, with SQ injections beginning on Day 0 and continuing every 14 (±3)days through Day 42.

Note: Extensions of the screening window to accommodate clinicallaboratory re-testing timeframes (excluding endoscopy with biopsy) arepermitted but are not allowed to exceed a total screening time of 16weeks.

Safety Assessments

Safety assessments include monitoring of AEs, clinical laboratory tests,vital signs measurements, physical examinations (including measurementof weight) and 12-lead ECG parameters. Demographic information andmedical and medication histories are obtained at the screening visit.

All safety assessments are recorded on the appropriate eCRF.

Demographic/Medical History

Demographic information, a complete medical history (which includessurgical history), and medication history is collected at the screeningvisit by appropriate site staff as delegated by the PI and reviewed andverified by a qualified licensed physician, physician's assistant, or anurse practitioner. The medical history is reviewed and recorded,including:

-   -   Date of birth    -   Sex    -   Race and ethnicity    -   Recent use of medication (30 days prior to enrollment for        non-Crohn's associated indications and lifetime recall for all        Crohn's associated medications)    -   CD past and ongoing treatments (recall past year)    -   CD-related surgical procedures and hospitalizations    -   History of respiratory, cardiovascular, renal, gastrointestinal,        hepatic, endocrine, hematological, neurological, psychiatric,        and other diseases; history of surgical procedures        Vital Signs

Vital signs, including systolic and diastolic blood pressure, pulse, andrespiration rate, are collected as shown in the Schedule of Assessments(see Table 13). Vital signs are within the ranges listed below duringScreening (Visit 1) and prior to enrollment at Visit 2:

-   -   Blood Pressure: 80/50 to 140/90 mm/Hg    -   Respiratory Rate: 8-20 breaths per minute    -   Pulse: 50-120 beats per minute    -   Temperature: 97.8° F. to 99.1° F. (36.5° C. to 37.3° C.)/average        98.6° F. (37° C.)

Vital signs are taken pre- and post-dose at Visits 2, 4, 6 and 8.Pre-dose vital signs are taken within 60 minutes before dosing.Post-dose vital signs are taken at least 60 minutes after dosing, priorto discharge. Additional blood pressure and pulse measurements areoptionally performed, as determined by the investigator, in order toensure appropriate monitoring of subject safety and accurate recordingof vital sign measurements. Any changes from baseline deemed clinicallysignificant by the investigator are recorded as AEs.

The same method of blood pressure measurement (auscultatory oroscillometric) is used and documented throughout the study for allsubjects. In addition, the conditions of vital signs measurements arecontrolled and as consistent as possible in order to minimizevariability of the readings. Measurements may be collected at acomfortable room temperature with little to no background noise, usingthe same (appropriately sized) cuff placed at the same location on thesame arm. The cuff has had a bladder length that is 80% and a width thatis at least 40% of arm circumference (a length-to-width ratio of 2:1).

The subject is asked to remove all clothing that covers the location ofcuff placement. The subject may not have exercised or consumed caffeine,alcohol, or nicotine within 30 minutes of collection. The subject may becomfortably seated, with the legs uncrossed, with feet flat on thefloor, and the back and arm supported, such that the middle of the cuffon the upper arm is at the level of the right atrium (the mid-point ofthe sternum). The subject is instructed to relax as much as possible forat least 5 minutes prior to collection. The subject may remain quietduring this time and throughout the measurement.

The bladder is deflated (calibrated for oscillometric method or manuallyby auscultatory method) at a rate of 2-3 mmHg/sec (and the first andlast audible sounds recorded as systolic and diastolic pressures) afterat least 5 minutes of rest.

The use of automated devices for measuring pulse is deemed acceptable,although, when done manually, pulse may be measured in thebrachial/radial artery for at least 30 seconds. When the timing of thesemeasurements coincides with a blood collection, blood pressure and pulsemay be obtained prior to the nominal time of the blood collection.

Physical Examination

A complete physical examination, including measurements of weight, isconducted by a qualified licensed physician, physician's assistant, or anurse practitioner at the screening visit and at Visits 2 (Day 0), 6(Day 28) and 10 (Day 56). Brief physical examinations, includingmeasurements of weight, are conducted at Visits 4 (Day 14), and 8 (Day42) (see Table 13).

The complete physical examination includes a review of the followingbody systems:

-   -   General appearance    -   Skin    -   Head, eyes, ears, nose, and throat    -   Spine/neck/thyroid    -   Musculoskeletal    -   Respiratory    -   Cardiovascular    -   Neurological    -   Abdomen (including liver and kidneys)

Any abnormalities or changes in intensity from baseline noted during thereview of body systems may be documented in the medical record andreported on the appropriate eCRF. If a new clinically significantabnormal finding is reported after the baseline examination, it isrequired to be captured as an AE and documented on the appropriate AEeCRF. In addition, resolution of any abnormal findings during the studyare noted in the medical record and eCRF if clinically significant.

The brief physical examination is to include a review of generalappearance, skin, head, eyes, ears, nose, and throat, abdomen, jointsand perianal area at a minimum, with other systems reviewed as medicallyneeded. Measurement of weight is also collected.

Subjects remove their shoes before measurements of weight (in kg) aretaken.

Electrocardiogram

A standard 12-lead ECG is conducted by appropriate site staff asdelegated by the PI and the results are reviewed and verified by the PIor a qualified licensed physician delegated by the PI as shown in theSchedule of Assessments (see Table 13).

The 12-lead ECG is performed after the subject has been supine forapproximately 5 minutes. All ECG recordings are identified with thesubject number, subject initials, date, and time of the recording andare included in the subject's study file.

The subject is asked to remove all clothing that covers the location oflead placement. The subject may not have exercised or consumed caffeine,alcohol, or nicotine within 30 minutes prior to collection.

In some cases, it may be appropriate to repeat abnormal ECGs to rule outimproper lead placement as contributing to the ECG abnormality. Leadsare placed in the same positions each time in order to achieve preciseECG recordings. One complete recording, including a 10-second rhythmstrip, may be taken at each time point. It may be immediately assessedas a valid recording and if not valid, it may be repeated. All ECGscollected are entered in the eCRF.

All ECGs are performed using the equipment supplied by the site. ECGrecordings are collected and a copy provided to the study sponsor.

The following parameters are recorded on the appropriate eCRF: heartrate, PR, respiration rate (RR), QRS, and QT interval; corrected QTintervals using both the Bazett (QTcB) and Fridericia (QTcF) formulasare also recorded. The investigator's assessment of the ECG tracing asnormal or abnormal is required to be documented, and if abnormal,his/her determination of whether the abnormality is clinicallysignificant is documented on the tracing and recorded in the eCRF.

All ECG values that, in the investigator's opinion, show clinicallyrelevant or pathological changes during or after termination of theinvestigational product are discussed with the medical monitor andreported as AEs and followed.

Clinical Laboratory Tests

Samples for the following clinical laboratory tests are collected at thetime points specified in the Schedule of Assessments (see Table 13).

TABLE 14 Clinical Laboratory Tests Test type Description HematologyHemoglobin, hematocrit, red blood cell count, red blood cell indices,mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration,mean corpuscular volume, platelet count (or estimate), and white bloodcell count including differential Serum chemistry Albumin, totalbilirubin, total protein, calcium, alkaline phosphatase, alanineaminotransferase, aspartate aminotransferase, gamma-glutamyltransferase, blood urea nitrogen, creatinine, creatine kinase, glucose,sodium, potassium, chloride, bicarbonate, lactate dehydrogenase, uricacid, eGFR, and C-reactive protein Other ADAs; LIGHT, cytokines (e.g.IL-1 beta, IL-6, IL-8, and TNF-alpha, and other exploratory cytokines),flow cytometry analysis of peripheral blood leukocytes and RNAsequencing Fecal chemistry Fecal calprotectin, C. difficile UrinalysispH, specific gravity, hemoglobin, white blood cells, red blood cells,glucose, casts, protein, ketones, epithelial cells, crystals, mucousthreads, bacteria, yeast, color, and appearance Serum & Urine β-hCG testFor women of childbearing potential only Urine drug screen Amphetamines,barbiturates, (screening visit only as benzodiazepines, cocaine,opiates, warranted) phencyclidine, cannabinoids, propoxyphene andmethadone Testing for previous Commercial test to detect the serumbiologies (for subjects concentrations of four of the common requiringwashout) biologic drugs and biosimilars (infliximab [Remicade],infliximad-adba [Renflexis], infliximab-dyyb [Inflectra], adalimumab[Humira], adalimumab-adbm [Cyltezo], adalimumab-atto [Amjevita]vedolizumab [Entyvio] and ustekinumab [Stelara]). Note: Testing notrequired for certolizumab pego1 (Cimzia) as there is no commerciallyavailable test to determine serum concentration.Sampled Blood Volume

The sampled blood volume that is taken from each subject is shown inTable 15.

TABLE 15 Blood Samples Taken from Each Subject Sample Number TotalVolume of Volume Assessment (mL) Samples (mL) Safety Hematology 2.0 612.0 Clinical chemistry 3.5 6 21.0 Anti-LIGHT monoclonal antibody 2.0 24.0 concentration and PK analysis (Pre-dose Day 0) Anti-LIGHT monoclonalantibody 2.0 9 18.0 concentration and PK analysis Anti-drug Antibodies(ADA) 2.0 5 10.0 Flow Cytometry 3.0 3 9.0 RNA sequencing 2.5 3 7.5Cytokines 3.5 3 10.5 LIGHT 2.0 3 6.0 Testing for previous biologies 6.01 6.0 Total mL — — 104.0Tuberculosis (TB) Testing

All subjects are screened for tuberculosis using QuantiFERON-TB Gold(QFT) blood test, or tuberculin skin reaction test (PPD skin test) atscreening. If the subject's PPD tine skin test is ≥5 mm, a chest x-rayis performed to rule out active or latent pulmonary TB infection.Subjects are excluded from the study if they have active or latent TB asdemonstrated by any of the following:

-   -   A positive QFT test result or a positive PPD skin test reaction        ≥10 mm.    -   Chest x-ray in which active or latent pulmonary TB cannot be        ruled out.        Genotyping for Decoy Receptor 3

Two milliliters of saliva for genotyping are collected in a designatedcollection vehicle according to the manufacturer's instructions.

Deoxyribonucleic acid (DNA) is isolated from the saliva samples fromeach study subject and then evaluated for genetic alteration in TNFRSF6Bencoding for the protein DcR3 or alterations in at least one DcR3network gene. All genotyping is performed in a CLIA certified laboratoryspecified in the laboratory manual(s) and guidance(s). Any remaining DNAsamples are stored for future biomarker studies.

LIGHT, Cytokines, RNA Sequencing and Flow Cytometry Exploratory Analyses

Blood samples are collected for exploratory analyses. Exploratoryanalyses include but are not limited to LIGHT biomarker, cytokines (e.g.IL-1 beta, IL-6, IL-8, TNF-alpha, and other exploratory cytokines),ribonucleic acid (RNA) sequencing; flow cytometry of peripheral bloodleukocytes at visits specified in Table 13.

Testing for Previous Biologics

An adequate wash-out period is required to avoid the potential forconfounding the effects related to dosing with the anti-LIGHT monoclonalantibody following the prior use of other biologics. In order to allowfor a shorter wash-out period, commercial tests available to identifythe serum concentrations of biologics are utilized. With the exceptionof Cimzia, a blood test is administered to subjects undergoing wash-out8 weeks after their last dose of biologic treatment to ensure that serumlevels of any previous biologic treatments are below the level ofdetection. If the serum concentration of biologic treatment isundetectable (i.e., below the level of quantification) according to therespective, CLIA-validated commercial test, then the subject is deemedto have completed the wash-out period and is permitted to proceed toVisit 2. If biologic treatment is detected, then the subject completes afull 12-week wash-out period prior to proceeding to Visit 2. Forsubjects receiving the biologic certolizumab pegol (Cimzia) a 12-weekwash-out is required.

Specimen Handling Requirements

The transmission of infectious agents may occur through contact withcontaminated needles, blood or blood products and/or laboratoryspecimens. Consequently, appropriate blood, body fluid and specimenprecautions are employed by all study personnel involved in thecollection and handling of specimens in both the clinic and laboratorysettings. Refer to current recommendations of the appropriateauthorities.

In addition to appropriate handling of subject samples, specificregulations exist regarding the shipment of biologic samples. Proceduresand regulations for the packaging and shipping of infectious samples areoutlined in the study Laboratory Manual(s). The investigator isresponsible for ensuring that all study samples that are to betransported to another location are appropriately packed and shippedaccording to the applicable regulations.

Evaluation of Laboratory Values

The normal ranges of values for the clinical safety laboratoryassessments are provided by the responsible laboratory and submitted tosponsor prior to the beginning of the study. They are regarded as thereference ranges upon which clinical decisions are made.

If a laboratory value is out of the reference range, it is notnecessarily clinically relevant, with some exceptions. The investigatoris required to evaluate the out-of-range values and record his/herassessment of their clinical relevance in the appropriate eCRF.

All laboratory values which, in the investigator's opinion, showclinically relevant or pathological changes during or after terminationof the treatment are discussed with the medical monitor and reported asAEs and followed.

Example 6.6—Adverse Events

Adverse Event Collection

The investigator is responsible for the detection and documentation ofevents meeting the criteria and definitions of an AE or SAE describedbelow. At each visit, the subject is allowed time to spontaneouslyreport any issues since the last visit or evaluation. At each visit, theinvestigator monitors, asks about, and/or evaluates any AEs usingnon-leading questions, such as:

-   -   “How are you feeling?”    -   “Have you experienced any issues since your last visit?”    -   “Have you taken any new medications since your last visit?”

Any clinically relevant observations made during each visit are alsoconsidered AEs.

Definition of Adverse Events, Period of Observation and Recording ofAdverse Events

An AE is defined as any untoward medical occurrence in a clinicalinvestigation subject administered a administered a pharmaceuticalproduct that does not necessarily have a causal relationship with theproduct. An AE can therefore be any unfavorable and unintended sign(including a new, clinically important abnormal laboratory finding),symptom, or disease, temporally associated with the product, whether ornot related to the product.

All AEs are collected from the time of the informed consent was signeduntil the final safety follow-up visit. This includes events occurringduring the screening phase of the study, regardless of whetherinvestigational product is administered. Where possible, a diagnosisrather than a list of symptoms is recorded. If a diagnosis has not beenmade, then each symptom is listed individually. All AEs are captured onthe appropriate AE eCRF and in source documents. In addition to AEs,unexpected benefits outside the investigational product indication arealso captured in the source documents and AE eCRF.

All AEs are followed to closure (ie, the subject's health has returnedto his/her baseline status or all variables have returned to normal),regardless of whether the subject is still participating in the study.Closure indicates that an outcome is reached, stabilization is achieved(the investigator does not expect any further improvement or worseningof the event), or the event is otherwise explained. When appropriate,medical tests and examinations are performed so that resolution of anevent(s) can be documented.

Severity of Adverse Events

The severity of AEs is recorded during the course of the event,including the start and stop dates for each change in severity. An eventthat changes in severity is captured as a new event. Worsening ofpre-treatment events after initiation of the investigational product arerecorded as new AEs. For example, if the subject experiences mild,intermittent headaches prior to dosing with investigational product andthe headache intensity increases to moderate after the first dose ofinvestigational product, a new AE of moderate intermittent headaches isrecorded in the source documents and eCRF.

The medical assessment of clinical severity of an AE is determined usingthe definitions outlined in Common Terminology Criteria for AdverseEvents (CTCAE), Version 4.0 (Published May 28, 2009 with Version 4.0.3on Jun. 14, 2010 by the US Department of Health and Human Services,National Institutes of Health, National Cancer Institute). Grade 1 isdefined as mild; asymptomatic or mild symptoms; or clinical ordiagnostic observations only; or intervention not indicated. Grade 2 isdefined as moderate; or minimal, local or non-invasive interventionindicated; or limiting age-appropriate instrumental activities of dailyliving (ADL). Grade 3 is defined as severe or medically significant butnot immediately life-threatening; or hospitalization or prolongation ofhospitalization indicated; or disabling; or limiting self-care ADL.Grade 4 is defined as life-threatening consequences; or urgentintervention indicated. Grade 5 is death related to AE.

Severity is a classification of intensity whereas an SAE is an AE thatmeets serious criteria. The above-referenced CTCAE document may bereferred to for full description of CTCAE terms and instrumental andself-care ADLs.

Relationship Categorization

A physician/investigator makes the assessment of relationship to theinvestigational product for each AE. The investigator decides whether,in his or her medical judgment, there is a reasonable possibility thatthe event could have been caused by the investigational product. Ifthere is no valid reason for suggesting a relationship, then the AE isclassified as “not related.” Otherwise, the AE is categorized accordingto the guidelines below. The causality assessment is documented in thesource document and the eCRF (Table 16).

TABLE 16 Assessment of Relationship to Investigational ProductRelationship Description Not Related Exposure to Investigational Product(IP) has not occurred. OR The administration of IP and the occurrence ofthe AE are not reasonably related in time. OR The AE is consideredlikely to be related to an etiology other than the use of the IP, thatis, there are no facts/evidence or arguments to suggest a causalrelationship to the IP. Possibly The administration of the IP and theoccurrence of Related the AE are reasonably related in time. AND The AEcould not be explained equally well by factors or causes other thanexposure to IP. Probably The administration of the IP and the occurrenceof Related the AE are reasonably related in time. AND The AE is morelikely explained by exposure to IP than by other factors or causes.Outcome at the Time of Last Observation

The outcome of an AE at the time of last observation is classified as:recovered/resolved; recovered/resolved with sequelae;recovering/resolving; not recovered/not resolved; fatal; or unknown.

Reporting of Serious Adverse Events

Initial and follow-up SAE reports may be completed by the investigatorand sent to the sponsor and the CRO within 24 hours of the firstawareness of a SAE. The investigator completes, signs and dates theappropriate SAE form and verifies the accuracy of the informationagainst corresponding source documents. No source documents are sentwith the SAE form. This SAE information (form) is sent to the CROpharmacovigilance department, with a copy to the sponsor's medicalmonitor by e-mail or fax.

Definition of Serious Adverse Event

An SAE is any untoward medical occurrence, whether considered to berelated to investigational product or not, that at any dose: results indeath; is life-threatening; requires inpatient hospitalization orprolongation of existing hospitalization; results in persistent orsignificant disability/incapacity; is a congenital anomaly; or is animportant medical event.

Note that the term “life-threatening” in the definition of “serious”refers to an event in which the subject was at risk of death at the timeof the event; it does not refer to an event which hypothetically mighthave caused death if it were more severe.

Note that inpatient hospitalization is defined as 24 hours in a hospitalor an overnight stay. An elective hospital admission to treat acondition present before exposure to the test drug, or a hospitaladmission for a diagnostic evaluation of an AE, does not qualify thecondition or event as an SAE. Further, an overnight stay in the hospitalthat is only due to transportation, organization, or accommodationproblems and without medical background does not need to be consideredan SAE.

Note that a congenital anomaly in an infant born to a mother who wasexposed to the investigational product during pregnancy is an SAE.However, a newly diagnosed pregnancy in a subject that has received aninvestigational product is not considered an SAE unless it is suspectedthat the investigational product interacted with a contraceptive methodand led to the pregnancy.

Note that medical and scientific judgment may be exercised in decidingwhether it is appropriate to consider other situations serious, such asimportant medical events that may not be immediately life-threatening orresult in death or hospitalization but may jeopardize the subject or mayrequire intervention to prevent one of the other outcomes listed in thedefinition above. Examples of such events are intensive treatment in anemergency room or at home for allergic bronchospasm, blood dyscrasias orconvulsions that do not result in hospitalization, or development ofdrug dependency or drug abuse.

Serious Adverse Event Collection Time Frames

All SAEs, regardless of the relationship to study, are collected fromthe time the subject signs the informed consent until the subject's lastvisit (office or telephone contact). The investigator or designeereports all SAEs promptly to the CRO and sponsor's medical monitorwithin 24 hours of first becoming aware of the event.

Any SAE(s), regardless of relationship to IP, discovered by theinvestigator at any interval after the study has completed is reportedto the CRO and sponsor's medical monitor within 24 hours of the firstawareness of the event.

Serious Adverse Event Onset and Resolution Dates

The onset date of the SAE is defined as the date the event meets seriouscriteria. The resolution date is the date the event no longer meetsserious criteria, the date symptoms resolve or the event is consideredchronic. In the case of hospitalization, the hospital admission anddischarge dates are considered respectively, the onset and resolutiondates of the SAE.

Any signs or symptoms experienced by the subject after signing theinformed consent form, or leading up to the onset date of the SAE orfollowing the resolution date of the SAE are recorded as AEs.

Fatal Outcome

An outcome of “fatal” may only be selected when the AE results in death.If more than 1 AE is possibly related to the subject's death, “fatal”outcome is indicated for each such AE.

Any AE that resulted in the subject's death was required have “fatal”checked as an outcome with the date of death recorded as the resolutiondate. Adverse events resulting in death may be reported within 24 hoursas SAEs, if not already reported as such.

For other AEs ongoing at the time of death that did not contribute tothe subject's death, the outcome is considered “not resolved” with noresolution date recorded.

Adverse Events of Special Interest and Adverse Drug Reactions

There are no events from research to date which qualify as an adverseevent of special interest. Adverse drug reactions observed in theanti-LIGHT monoclonal antibody pre-clinical study include injection sitereactions.

Adverse drug reactions with other biologic agents include: potential forincreased infection (including opportunistic infections such astuberculosis); hypersensitivity reactions (including anaphylaxis);immunogenicity; malignancy; impaired immunization; or CD exacerbation.

Pregnancy

All females of childbearing potential who participate in the study arecounseled on the need to practice adequate birth control and on theimportance of avoiding pregnancy during study participation. Studyparticipants are instructed to contact the investigator or study staffimmediately if pregnancy occurs or is suspected.

Pregnancy testing is also conducted prior to administration ofinvestigational product on every female of childbearing potential. Anyfemale who is found to be pregnant at the screening visit is excludedfrom the study and considered a screening failure.

The investigator reports the pregnancy of any female (study participantor female partner of male study participant) who becomes pregnant duringinvestigational product treatment or within 7 days of discontinuing theinvestigational product. The pregnancy is reported within 24 hours ofawareness to the CRO. The investigator contacts the designatedindividual(s) who receive SAE notification and record informationrelated to the pregnancy on the designated form provided by the sponsoror its designee.

Early termination visit assessments are conducted as soon as possibleafter learning of the pregnancy. The investigator is also responsiblefor following the pregnancy until delivery or termination. Thesefindings are reported on the Exposure in Utero form/other designatedform and forwarded to the designated individual(s). The event meets theSAE criterion only if it results in a spontaneous abortion or acongenital anomaly.

Example 6.7—Abuse, Misuse, Overdose, and Medication Error

Abuse, misuse, overdose or medication error involving theinvestigational product, as defined below, is reported to the sponsorusing the SAE reporting procedures whether or not they result in an AEor SAE. The 24-hour reporting period from time of first awareness doesnot apply to an abuse, misuse, overdose, or medication error event(s)unless the abuse, misuse, overdose, or medication error event results ina SAE.

TABLE 17 Definitions of Abuse, Misuse, Overdose, and Medication ErrorCategory Definition Abuse Persistent or sporadic intentional intake ofIP when used for a non- medical purpose (for example, to get high, forpotential psychoactive effects) in a manner that would be detrimental tothe individual and/or society Misuse Intentional use of IP other than asdirected or indicated at any dose. This includes where IP is not used asdirected at the dose prescribed in the protocol. Overdose Intentional orunintentional intake of a dose of IP exceeding the dose prescribed tothe subject as part of the study. Medication Error made in prescribing,dispensing, administration Error and/or use of IP. A medication error isreportable to the sponsor or its designee if it involves: Administrationand/or use of an unassigned treatment (for example, incorrect IP kitused by subject) Administration and/or use of expired IP. IP =investigational product Note that an abuse, misuse, overdose ormedication error event can meet more than 1 category. Missing doses arenot considered medication error events and do not need to be reported.

Example 6.8—Anti-LIGHT Monoclonal Antibody Concentration,Pharmacokinetic and Anti-Drug Antibody Assessments

Pharmacokinetics are calculated from the plasma concentrations of theanti-LIGHT monoclonal antibody.

Blood Sample Collection and Analysis

Blood samples are collected within 60 minutes prior to theadministration of the anti-LIGHT monoclonal antibody on Days 0, 14, 28and 42 (Visits 2, 4, 6, and 8 respectively) and at any time on Days 7,21, 35, 49, 56, and 84 (Visits 3, 5, 7, 9, 10, and 11) and processed toplasma. Time of PK samples is recorded in the electronic CRF. A total of1.0 mL plasma (0.5 mL per PK and ADA sample) is collected from eachsubject to measure plasma concentrations of the anti-LIGHT monoclonalantibody or ADA samples. Pharmacokinetic and ADA samples are processedaccording to the methods and directions set forward in the LaboratoryManual(s) and guidance(s).

Pharmacokinetic and ADA plasma sample analysis is performed by aspecified laboratory according to their SOPs using a validatedenzyme-linked immunosorbent assay (ELISA).

Example 6.9—Assessment of Efficacy

Crohn's Disease Activity Index

The CDAI is completed by a qualified licensed physician, physician'sassistant, or a nurse practitioner at the times shown in the Schedule ofAssessments (see Table 13). Site personnel conducting the CDAI deriveindividual and total scores, with standard weights calculated asfollows: standard weight for men=(height in m)2×22.1; standard weightfor women=(height in m)2×20.8.

Information on abdominal pain and frequency of loose and watery stoolsis taken from subject diary information. The same individual comples theCDAI for a given subject throughout the study, whenever possible.

The CDAI was developed by the National Cooperative Crohn's Disease Studygroup and published in 1976 by Best et al. (1976) (Best et al.,Development of a Crohn's disease activity index, National CooperativeCrohn's Disease Study, Gastroenterol., 1976:70; 439-44) to determinevariables that best predicted disease activity. A total of 8 items wereidentified (abdominal pain, number of liquid stools, general well-being,extraintestinal complication, use of antidiarrheal drugs, abdominalmass, hematocrit, and body weight). Each item is scored on individualparameter criteria. Total CDAI scores can range from 0 to approximately600 with higher scores indicating more active disease. The CDAI has beenthe most frequently used efficacy scale for interventional studies in CD(Sostegni et al., Review: Crohn's disease: monitoring disease activity,Aliment Pharmacolo Ther., 2003:17 (Suppl. 2) 11-17).

Endoscopy with Biology and Histology

All subjects who enroll in the study undergo an endoscopy with biopsy atscreening and again at Day 56 (Visit 10) or at early termination.Screening endoscopies are optionally performed either as a stand-aloneendoscopy for purposes of this protocol or as a clinically-requiredendoscopy, provided consenting procedures for this study are completedprior to endoscopy.

Endoscopy evaluation uses the SES-CD. The SES-CD is a simple,easy-to-use endoscopic scoring system developed specifically for CD. Itassesses 4 variables: size of ulcers, percentage of ulcerated surface,percentage of affected surface and the presence of narrowing across 4categories per variable on a scale of 0 to 3 (Daperno M, D'Haens G, VanAssche G et al. Development and validation of a new, simplifiedendoscopic activity score for Crohn's disease: the SES-CD. GastroinestEndosc. 2004 October; 60(4):505-12). Biopsies taken at screening areassessed for histological confirmation of disease. Each subject hasscreening and Visit 10/ET biopsy samples retained for evaluation ofexploratory parameters (which may include but is not limited to DcR3,LIGHT, HVEM and LTβR) which could optionally occur after studycompletion.

Patient-Reported Assessment of Well-being, Abdominal Pain and StoolFrequency

All subjects who enroll in the study report their daily assessment ofwell-being, abdominal pain and stool frequency including loose and/orwatery stools via a diary (electronic or hard copy). Abdominal pain isassessed on a scale of 0 to 3 with higher values indicating greater painseverity. The stool frequency including number of loose and/or waterystools per day, equivalent to a score of a 6 or 7 on the Bristol StoolScale, is recorded.

Loose stools are described as fluffy pieces with ragged edges, a mushystool. Watery stools are described as watery, no solid pieces (O'Donnellet al., Detection of pseudodiarrhoea by simple clinical assessment ofintestinal transit rate, Br. Med. J. 1990; 300:439-40).

Quality of Life Assessment—Inflammatory Bowel Disease Questionnaire

The IBD-Q is a 32 item questionnaire validated to measure quality oflife in Crohn's Disease. The IBD-Q assesses the dimensions of bowelfunction, emotional status, systemic symptoms and social function(Guyatt et al., A new measure of health status for clinical trials ininflammatory bowel disease, Gastroenterol., 1989:96; 804-10). The IBD-Qis completed by all subjects at screening (Visit 1), before dosing(Visit 2), and at the end of the open-label treatment period or earlytermination (Visit 10/ET) (see Table 13).

Example 6.10—Statistics

Pharmacokinetic, efficacy and quality of life data is summarized withtraditional descriptive statistics. Continuous variables are summarizedwith N, mean, standard deviation, and range. Categorical variables aresummarized by frequencies and percentages.

No formal inferential analyses are planned.

The Safety Population includes all subjects who enroll in the study andreceive any amount of investigational product. The PharmacokineticPopulation includes all subjects who receive their assigned dose of theanti-LIGHT monoclonal antibody and for whom the anti-LIGHT monoclonalantibody plasma concentration data is available. The Efficacy Populationincludes all subjects who have a baseline and at least 1 post-baselineefficacy score.

Analyses of efficacy focus on the Efficacy Population. Results ofendpoints (e.g., CDAI, SES-CD, abdominal pain and loose/watery stoolfrequency, and IBD-Q) are summarized by visit for each cohort, both asraw scores and the change from baseline value. CDAI individual and totalscores are derived programmatically using recorded data from patientdiary, responses to CDAI questions, laboratory tests, and physical examresults.

Quantitative endoscopy and biopsy results are also summarized for boththe Baseline and End of study visits.

Analyses of safety data are focused on the Safety Population.

Safety variables include treatment-emergent AEs (TEAEs), clinicallaboratory results, vital signs measurements, ECG results, and physicalexamination findings.

Adverse events are coded using Medical Dictionary for Regulatory Affairs(MedDRA) version 20. TEAEs are defined as any AE having first onset orworsening in severity after the first administration of IP. TEAEs areclassified by system organ class (SOC) and preferred term and summarizedby the number of subjects reporting each event for each cohort andoverall. Similar summaries are produced for SAEs, AEs leading todiscontinuation, and AEs with at least a possible relationship to theinvestigational product. The intensity of AEs and the relationship tothe investigational product are also summarized for each SOC andpreferred term.

For clinical laboratory tests, descriptive summaries of actual(absolute) values and change from baseline values are presented bycohort for each study visit. The number of subjects with clinicallaboratory values below, within, or above normal ranges at each studyvisit are tabulated (shift tables) for each clinical laboratory test bycohort.

Vital signs (systolic and diastolic blood pressure, pulse, andrespiratory rate) and ECG results are summarized by visit and cohortusing appropriate descriptive statistics. The number and percentage ofsubjects with abnormal ECG findings is summarized by cohort for eachstudy visit.

Data in this open-label study is monitored continually.

This is the first use of the anti-LIGHT monoclonal antibody in theintended population of patients with CD resistant to anti-TNFαmonoclonal antibodies. The sample size of the study was based onfeasibility.

Example 7

Presented herein in Example 7 are certain results of the study conductedaccording to the methods as described in Example 6.

Three timepoint serum samples were obtained from the two patients whohave completed anti-LIGHT antibody treatment in the study described inExample 6. Healthy adult donors (n=30 controls) had an averagepre-treatment free LIGHT level of 202 pg/mL. The first patient had anelevated plasma pre-treatment free LIGHT level of 455 pg/mL. Aftertreatment with the anti-LIGHT antibody (having a VH of SEQ ID NO. 8, anda VL of SEQ ID NO: 9) subcutaneous (SQ) every (q) 2 weeks, the subject'splasma free LIGHT levels were found to be in the normal range;free-LIGHT levels were 15 and 24 pg/mL on days 28 and 56, respectively.The second patient had an elevated plasma pre-treatment free LIGHT levelof 193 pg/mL. After treatment with the anti-LIGHT antibody subcutaneous(SQ) every (q) 2 weeks, the subject's plasma free LIGHT levels werefound to be in the normal range; free-LIGHT levels were 42 and 29 pg/mLon days 28 and 56, respectively. These data show that in the outpatientsetting even relatively low doses of the anti-LIGHT antibody candecrease plasma free LIGHT levels, and that the free LIGHT levels canstay decreased.

Moreover, the first patient's clinical improvement was seen to correlatewith this reduction. Simple Endoscopic Score for Crohn's Disease(SES-CD) the first patient's score decreased with treatment. An SES-CDScore of: 0-2 means remission; 3-6 means mild; 7-15 means moderate;and >15 means severe. The first patient's SES-CD score was 11 atscreening and was 4 at day 56.

Example 8

LIGHT testing using the free LIGHT assay described herein was performedon samples from ARDS patients and compared to healthy donor LIGHTlevels. The preliminary data shows elevated LIGHT levels in the ARDSpopulation compared to healthy donor levels. A box plot generated onthis data is shown in FIG. 11 .

The following Table 18 provides the sequences referred to in thisapplication.

TABLE 18 Table of Sequences SEQ ID NO DESCRIPTION SEQUENCE  1 Human DcR3MRALEGPGLS LLCLVLALPA LLPVPAVRGV amino acid AETPTYPWRD sequenceAETGERLVCA QCPPGTFVQR PCRRDSPTTC GPCPPRHYTQ FWNYLERCRY CNVLCGEREEEARACHATHN RACRCRTGFF AHAGFCLEHA SCPPGAGVIA PGTPSQNTQC QPCPPGTFSASSSSSEQCQP HRNCTALGLA LNVPGSSSHD TLCTSCTGFP LSTRVPGAEE CERAVIDFVAFQDISIKRLQ RLLQALEAPE GWGPTPRAGR AALQLKLRRR LTELLGAQDG ALLVRLLQALRVARMPGLER SVRERFLPVH  2 Heavy Chain GYNWH (HC) CDR1 antibody F19  3HC CDR2 EITHSGSTNYNPSLKS antibody F19  4 HC CDR3 EIAVAGTGYYGMDVantibody F19  5 LC CDR1 RASQGINSAFA antibody F19  6 LC CDR2 DASSLESantibody F19  7 LC CDR3 QQFNSYPLT antibody F19  8 Heavy chainQVQLQQWGAG LLKPSETLSL TCAVYGGSFS variable GYNWHWIRQP PGKGLEWIGE ITHSGSTNYN region PSLKSRVTIS VDTSKNQFSL KLSSVTAADTantibody F19 AVYYCVREIA VAGTGYYGMD VWGQGTTVTVSSASTKGPSV FPLAPCSRST SESTAALGCL VKDYFPEPVT VSWNSGALTS GVHTFPAVLQSSGLYSLSSV VTVPSSSLGT KTYTCNVDHK PSNTKVDKRV ESKYGPPCPP CPAPEFEGGPSVFLFPPKPK DTLMISRTPE VTCVVVDVSQ EDPEVQFNWY VDGVEVHNAK TKPREEQFNSTYRVVSVLTV LHQDWLNGKE YKCKVSNKGL PSSIEKTISK AKGQPREPQV YTLPPSQEEMTKNQVSLTCL VKGFYPSDIA VEWESNGQPE NNYKTTPPVL DSDGSFFLYS RLTVDKSRWQEGNVFSCSVM HEALHNHYTQ KSLSLSLG  9 Light chainAIQLTQSPSS LSASVGDRVT ITCRASQGIN variable SAFAWYQQKP GKAPKLLIYD ASSLESGVPS region RFSGSGSGTD FTLTISSLQP EDFATYYCQQantibody F19 FNSYPLTFGG GTKVEIKRTV AAPSVFIFPPSDEQLKSGTA SVVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLSSTLTLSKADYEKHK VYACEVTHQG LSSPVTKSFN  RGEC 10 Alternative  RASRGINSAFALC CDR1  antibody F19 11 Alternative  DASSLES LC CDR2  antibody F19 12Alternative  QQFNSYPLT LC CDR3  antibody F19 13 Alternative  RVSQGISSYLNLC CDR1  antibody F19 14 Alternative  SASNLQS LC CDR2  antibody F19 15Alternative  ARTNAPPT LC CDR3  antibody F19 16 Alternative  RMSQGISSYLALC CDR1  antibody F19 17 Alternative  AASTLQS LC CDR2  antibody F19 18Alternative  QQYYSFPYT LC CDR3  antibody F19 19 Alternative  RASQGVSSYLALC CDR1  antibody F19 20 Alternative  DASNRAT LC CDR2  antibody F19 21Alternative  QQRSNWHP LC CDR3  antibody F19 22 HC CDR1 RFNMN antibody E123 HC CDR2 YISSSSYTIYYADSVKG antibody E1 24 HC CDR3 SIAAFDY antibody E125 LC CDR1 RASQGISSALA antibody E1 26 LC CDR2 DASSLES antibody E1 27LC CDR3 QQFNSYRT antibody E1 28 Alternative  RASQSVSSSYLT LC CDR1 antibody E1 29 Alternative  GASSRAT LC CDR2  antibody E1 30 Alternative QQYGSSMYT LC CDR3  antibody E1 31 Alternative  RASQSVSSSYLA LC CDR1 antibody E1 32 Alternative  GASNRAT LC CDR2  antibody E1 33 Alternative QQYGSSPWT LC CDR3  antibody E1 34 HC CDR1 NAWMS antibody E13 35 HC CDR2RIKSKIDGGTTDYAAPVKG antibody E13 36 HC CDR3 AMAGAFGF antibody E13 37LC CDR1 RASQSVSSSYLA antibody E13 38 LC CDR2 GASSRAT antibody E13 39LC CDR3 QQYGSSPMYT antibody E13 40 HC CDR1 SGGYYWS antibody E63 41HC CDR2 YIYYSGSTNYNPSLKS antibody E63 42 HC CDR3 WITMFRGVGFDPantibody E63 43 LC CDR1 RASQSIGSSLH antibody E63 44 LC CDR2 YASQSFSantibody E63 45 LC CDR3 RQSSSLPLT antibody E63 46 HC CDR1 GYYWNantibody F23 47 HC CDR2 EINQYNPSLKS antibody F23 48 HC CDR3EIAIADKGYYGLDV antibody F23 49 LC CDR1 RASQGISSALA antibody F23 50LC CDR2 DASSLES antibody F23 51 LC CDR3 QQFNSYPLT antibody F23 52HC CDR1 SYYIH 53 HC CDR2 PGSDITKYNEKFKG 54 HC CDR3 GISTYSAMDF 55 LC CDR1KASQDVGTAVA 56 LC CDR2 WASTRHT 57 LC CDR3 QQYSSYPLT 58 HC variableQVQLVQSGAE VKKPGASVKV SCKASGYTFT region SYYIHWVRQA PGQRLEWMGW IFPGSDITKYNEKFKGRVTI TRDTSASTAY MELSSLRSED TAVYYCARED YGISTYSAMD FWGQGTLVTV  SS 59LC variable DIQLTQSPSF LSASVGDRVT ITCKASQDVG regionTAVAWYQQKP GKAPKLLIYW ASTRHTGVPS RFSGSGSGTE FTLTISSLQP EDFATYYCQQYSSYPLTFGQ GTKVEIKR 60 HC CDR1 of HFDIN 18E04 61 HC CDR2 ofWMNPDSDNTDYAQEFQG 18E04 62 HC CDR3 of GGTTLDY 18E04 63 LC CDR1 ofSGDALPKKYAY 18E04 64 LC CDR2 of EDSKRPS 18E04 65 LC CDR3 of YSTDSSDNHVI18E04 66 HC CDR1 of DYYMS 98C07 67 HC CDR2 of YISRSSFIYYSESVKG 98C07 68HC CDR3 of WELSPFDY 98C07 69 LC CDR1 of RASQGISNYLA 98C07 70 LC CDR2 ofAASSLQS 98C07 71 LC CDR3 of QQYNTYPFT 98C07 72 HC CDR1 of YYGIS 1C02 73HC CDR2 of WISANSGNTNYAQKFQG 1C02 74 HC CDR3 of GGVAVLEY 1C02 75LC CDR1 of WASQGISSYLA 1C02 76 LC CDR2 of VASTLQS 1C02 77 LC CDR3 ofQQLKIYPLT 1C02 78 HC CDR1 of DYYMN 1C06 79 HC CDR2 of DISSRDNTIYYADSVKG1C06 80 HC CDR3 of ARERGFGDYFGMDV 1C06 81 LC CDR1 of RASQDISSALA 1C06 82LC CDR2 of DASSLES 1C06 83 LC CDR3 of QQFNTYPLT 1C06

What is claimed is:
 1. A method of treating acute respiratory distress syndrome (ARDS) or acute lung injury (ALI) associated with COVID-19 comprising administering to a human subject in need thereof an effective amount of an anti-LIGHT antibody comprising a heavy chain and a light chain variable region comprising a CDR-H1 having the sequence of SEQ ID NO: 2, CDR-H2 having the sequence of SEQ ID NO: 3, CDR-H3 having the sequence of SEQ ID NO: 4, CDR-L1 having the sequence of SEQ ID NO: 5, CDR-L2 having the sequence of SEQ ID NO: 6, and CDR-L3 having the sequence of SEQ ID NO:
 7. 2. The method of claim 1, wherein the subject has acute respiratory distress syndrome (ARDS).
 3. The method of claim 1, wherein the subject has acute lung injury (ALI).
 4. The method of claim 1, wherein the subject has pneumonia.
 5. The method of claim 1, wherein administration of the anti-LIGHT antibody reduces serum free-LIGHT levels in the subject.
 6. The method of claim 1, wherein a single dose of about 16 mg/kg of the anti-LIGHT antibody is administered.
 7. The method of claim 1, wherein the subject has received, or is currently receiving, an anti-COVID-19 therapy comprising a corticosteroid, hydroxychloroquine, and/or remdesivir.
 8. The method of claim 1 wherein administration of the anti-LIGHT antibody reduces the subject's risk of mortality by equal to or greater than 50% at 28 days and/or 60 days after administration.
 9. The method of claim 8, where the subject is 60 years of age or older.
 10. The method of claim 1, wherein administration of the anti-LIGHT antibody reduces the subject's risk of respiratory failure. 